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Qingxiang Amy Sang - One of the best experts on this subject based on the ideXlab platform.

  • The Anti-inflammatory Role of Endometase/Matrilysin-2 in Human Prostate Cancer Cells
    2016
    Co-Authors: Zahraa I Khamis, Diego A. R. Zorio, W. K. Chung, Qingxiang Amy Sang
    Abstract:

    licenses/by-nc-nd/3.0/). Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited. Received: 2013.01.01; Accepted: 2013.02.07; Published: 2013.03.20 Human endometase/matrilysin-2/Matrix Metalloproteinase-26 (MMP-26) is an endopeptidase mostly produced by human carcinoma cells. While MMPs are thought to regulate the dynamics of extracellular Matrix turnover, new evidence shows that these enzymes may play a critical regu-latory role in inflammation. To investigate the role of MMP-26 in inflammation, three different variants of androgen repressed human prostate cancer (ARCaP) cells were investigated in the study: parental, MMP-26 sense cDNA-transfected, and MMP-26 antisense cDNA-transfected ARCaP cells. Protein lysates and RNA from control and genetically modified cells were analyzed by Western blotting and real-time reverse transcription polymerase chain reaction on arrays of genes critical to the inflammatory response. In comparison to parental controls, up-regulation o

  • evidence for a proapoptotic role of Matrix Metalloproteinase 26 in human prostate cancer cells and tissues
    Journal of Cancer, 2016
    Co-Authors: Zahraa I Khamis, Yangao Man, Kenneth A Iczkowski, Mayassa J Boudargham, Qingxiang Amy Sang
    Abstract:

    Matrix Metalloproteinases (MMPs) play intricate roles in cancer progression; some promote invasion and angiogenesis while others suppress tumor growth. For example, human MMP-26/endometase/matrilysin-2 was reported to be either protective or pro-tumorigenic. Our previous reports suggested pro-invasion and anti-inflammation properties in prostate cancer. Here, we provide evidence for a protective role of MMP-26 in the prostate. MMP-26 expression levels in androgen-repressed human prostate cancer (ARCaP) cells, transfected with sense or anti-sense MMP-26 cDNA, are directly correlated with those of the pro-apoptotic marker Bax. Immunohistochemical staining of prostate cancer tissue samples shows similar protein expression patterns, correlating the expression levels of MMP-26 and Bax in benign, neoplastic, and invasive prostate cancer tissues. The MMP-26 protein levels were upregulated in high grade prostate intraepithelial neoplasia (HGPIN) and decreased during the course of disease progression. Further analysis using an indirect terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay showed that many tumor cells expressing MMP-26 were undergoing apoptosis. This study showed that the high level of MMP-26 expression is positively correlated with the presence of apoptotic cells. This pro-apoptotic role of MMP-26 in human prostate cancer cells and tissues may enhance our understanding of the paradoxical roles of MMP-26 in tumor invasion and progression.

  • efficient soluble expression of secreted Matrix Metalloproteinase 26 in brevibacillus choshinensis
    Protein Expression and Purification, 2013
    Co-Authors: Weiguo Liang, Qingxiang Amy Sang, Zhiyong Wang, Zhongyuan Wang, Mark D Roycik, Hongyu Xiang, Xuexun Fang
    Abstract:

    Matrix Metalloproteinase 26 (MMP-26) is a novel member of the Matrix Metalloproteinase family with minimal domain constitution and unknown physiological function. The three-dimensional (3D) structure of the enzyme also remains to be deciphered. Previous studies show that MMP-26 may be expressed in Escherichia coli (E. coli) as inclusion bodies and re-natured with catalytic activity. However, the low re-naturation rate of this method limits its usage in structural studies. In this paper, we tried to clone, express and purify the pro form and catalytic form of MMP-26 (ProMMP-26 and CatMMP-26) in several widely used expression vectors and express the recombinant MMP-26 proteins in E. coli cells. These constructs resulted in insoluble expressions or soluble expressions of MMP-26 with little catalytic activity. We then used Brevibacillus choshinensis (B. choshinensis) as the host system for the soluble and active expression of MMP-26. The enzyme was secreted in soluble form in the supernatant of cell culture medium and purified via a two-step purification process that included Ni(2+) affinity chromatography followed by gel filtration. The yields of purified ProMMP-26 and CatMMP-26 were 12 and 18mg/L, respectively, with high purity and homogeneity. Both ProMMP-26 and CatMMP-26 showed gelatin zymography activity and the purified CatMMP-26 had high enzymatic activity against DQ-gelatin substrate. The large-scale soluble and active protein production for future structural studies of MMP-26 is thus feasible using the B. choshinensis host system.

  • Matrix Metalloproteinase 26 matrilysin 2 homo sapiens
    Handbook of Proteolytic Enzymes, 2013
    Co-Authors: Qingxiang Amy Sang
    Abstract:

    The third edition of the Handbook of Proteolytic Enzymes aims to be a comprehensive reference work for the enzymes that cleave proteins and peptides, and contains over 850 chapters. Each chapter is organized into sections describing the name and history, activity and specificity, structural chemistry, preparation, biological aspects, and distinguishing features for a specific peptidase. The subject of Chapter 162 is Matrix Metalloproteinase-26/Matrilysin 2.

  • Protein Signatures in Human MDA-MB-231 Breast Cancer Cells Indicating a More Invasive Phenotype Following Knockdown of Human Endometase/Matrilysin-2 by siRNA
    Ivyspring International Publisher, 2011
    Co-Authors: Seakwoo Lee, Doris Terry, Douglas R. Hurst, Danny R. Welch, Qingxiang Amy Sang
    Abstract:

    Human Matrix Metalloproteinase-26 (MMP-26/endometase/matrilysin-2) is a putative biomarker for carcinomas of breast, prostate, and other cancers of epithelial origin. MMP-26 expression was silenced using small interfering RNA (siRNA) in the human breast cancer cell line MDA-MB-231. Immunological and proteomics approaches, including two-dimensional gel electrophoresis and Matrix assisted laser desorption/ionization time-of-flight mass spectrometry, were employed to identify differential protein expression in MMP-26 knockdown cells. A comparison of the protein expression profiles of control and MMP-26 knockdown cells revealed nine differentially regulated proteins. Five of the proteins (heat shock protein 90, glucose-regulated protein 78 (GRP78), annexin V, tropomyosin, and peroxiredoxin II) were up-regulated, while alpha-tubulin, cystatin SA-III, breast cancer metastasis suppressor 1 (BRMS1) and beta-actin were down-regulated. This decrease of BRMS1 expression is concomitant with an increase of invasion through Matrix-coated membranes. These results suggest an important role for MMP-26 in the regulation of proteins involved in invasive and metastatic breast cancers.

Yunge Zhao - One of the best experts on this subject based on the ideXlab platform.

  • expression of Matrix Metalloproteinase 26 in multiple human cancer tissues and smooth muscle cells
    Chinese Journal of Cancer, 2009
    Co-Authors: Yunge Zhao, Aizhen Xiao, Yangao Man, Qingxiang Amy Sang
    Abstract:

    BACKGROUND AND OBJECTIVE Elevated expression of Matrix Metalloproteinases (MMPs) has been found in multiple carcinoma tissues. MMP-26 is highly expressed in prostate and breast cancer tissues, and promotes the invasion of human prostate cancer cells not only through the cleavage of fibronectin and type IV collagen but also by the activation of pro-MMP-9, a powerful gelatinase. This study was to present a comprehensive protein expression profile of MMP-26 in multiple human cancer tissues. METHODS The protein expression pattern of MMP-26 was examined using immunohistochemistry and multiple-tissue microarray. MMP-26 mRNA expression in coronary artery smooth muscle cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS The expression of MMP-26 in breast, colon, lung, brain, head and neck, prostate cancer, and melanoma tissues was significantly elevated when compared with parallel normal tissues (P 0.05). MMP-26 was also detected to express in gastric, rectal, thyroid, esophageal, and pancreatic cancers. MMP-26 protein was expressed in smooth muscle cells of the prostate and associated blood vessels. MMP-26 mRNA was also detected to express in human coronary artery smooth muscle cells. CONCLUSIONS MMP-26 expression may be associated with multiple human carcinomas, and it may serve as a molecular marker for the early diagnosis of these carcinomas. MMP-26 may also contribute to smooth muscle function in the human prostate and cardiovascular system.

  • spatio temporal expression of Matrix Metalloproteinase 26 in human placental trophoblasts and fetal red cells during normal placentation
    Biology of Reproduction, 2005
    Co-Authors: Wei Qiu, Yunge Zhao, Qingxiang Amy Sang, Suxia Bai, Meirong Zhao, Yanling Wang
    Abstract:

    The processes of implantation and placentation involve the degradation and remodeling of extracellular Matrix, cellular proliferation, apoptosis, and differentiation. Evidence indicates that members of the Matrix Metalloproteinase (MMP) family play crucial roles in these processes. In the present study, we identified the expression and localization of MMP26/endometase/ matrilysin-2 in human placentae at different stages of gestation using methods of reverse transcriptase-polymerase chain reaction, in situ hybridization, and immunohistochemistry. MMP26 was widely localized to villous cytotrophoblast cells, syncytiotrophoblast cells, and to column trophoblasts during early pregnancy. The mRNA and protein level of MMP26 in chorionic villi was highest at Weeks 6-7, and decreased thereafter, reaching its lowest level at the second trimester. The mRNA level was significantly up-regulated in term placenta, while the immunoreactivity remained undetectable. Notably, intense expression of MMP26 was found in fetal nucleated red cells inside the villous capillaries during gestational Weeks 6-9. Strong expression of MMP26 mRNA was also demonstrated in fetal red cells isolated from the whole blood of fetuses at midpregnancy. The expression patterns of MMP26 in human placenta suggests complicated roles for MMP26 during the processes of placentation and hematopoiesis, perhaps working in concert with other members of the MMP family, such as MMP9.

  • promoting breast cancer cell invasion by Matrix Metalloproteinase 26 in a novel three dimensional pva sponge culture system
    2004
    Co-Authors: Yunge Zhao
    Abstract:

    Abstract : Cells are grown in 3-dimensional environments under physiological and pathological condition. While most of the experiments were done using the monolayer cell culture. Accumulating results showed that the expression patterns of genes and proteins are distinctly different in three-dimensional (3D) cultures when compared with the monolayer (2D) cultures commonly employed. Therefore, we created a novel Polyvinyl alcohol (PVA)3D cell culture system to study human breast cancer cell progression and cancer cell-stromal cell interaction. The results showed that both breast cancer cell (MDA-MB-23l)and human fibroblast cell (HFL-l)attachment varied with the different extra cellular Matrix components. When the MDA-MB-23l cell invaded into the bottom layer, which contained HFL-l cell in the PVA 3D cell culture, the cells formed breast cancer tissue-like structure. Furthermore, the MDA-MB-23l cell could be easily identified immunohistochemistry using proper epithelial marker. Therefore, This 3D culture system may be a useful medol to study cancer cell-stromal cell interaction and cancer cells invasion. The role of the novel MMP-26 in breast cancer invasion using this 3D cell culture system is under investigation.

  • activation of pro gelatinase b by endometase matrilysin 2 promotes invasion of human prostate cancer cells
    Journal of Biological Chemistry, 2003
    Co-Authors: Yunge Zhao, Hyun I Park, Aizhen Xiao, Robert G Newcomer, Tiebang Kang, Leland W K Chung, Mark G Swanson, Haiyen E Zhau, John Kurhanewicz, Qingxiang Amy Sang
    Abstract:

    Abstract This work has explored a putative biochemical mechanism by which endometase/matrilysin-2/Matrix Metalloproteinase-26 (MMP-26) may promote human prostate cancer cell invasion. Here, we showed that the levels of MMP-26 protein in human prostate carcinomas from multiple patients were significantly higher than those in prostatitis, benign prostate hyperplasia, and normal prostate glandular tissues. The role of MMP-26 in prostate cancer progression is unknown. MMP-26 was capable of activating pro-MMP-9 by cleavage at the Ala93–Met94 site of the prepro-enzyme. This activation proceeded in a time- and dose-dependent manner, facilitating the efficient cleavage of fibronectin by MMP-9. The activated MMP-9 products generated by MMP-26 appeared more stable than those cleaved by MMP-7 under the conditions tested. To investigate the contribution of MMP-26 to cancer cell invasion via the activation of MMP-9, highly invasive and metastatic human prostate carcinoma cells, androgen-repressed prostate cancer (ARCaP) cells were selected as a working model. ARCaP cells express both MMP-26 and MMP-9. Specific anti-MMP-26 and anti-MMP-9 functional blocking antibodies both reduced the invasiveness of ARCaP cells across fibronectin or type IV collagen. Furthermore, the introduction of MMP-26antisense cDNA into ARCaP cells significantly reduced the MMP-26 protein level in these cells and strongly suppressed the invasiveness of ARCaP cells. Double immunofluorescence staining and confocal laser scanning microscopic images revealed that MMP-26 and MMP-9 were co-localized in parental and MMP-26 sense-transfected ARCaP cells. Moreover, MMP-26 and MMP-9 proteins were both expressed in the same human prostate carcinoma tissue samples examined. These results indicate that MMP-26 may be a physiological and pathological activator of pro-MMP-9.

  • identification and specific expression of Matrix Metalloproteinase 26 in rhesus monkey endometrium during early pregnancy
    Molecular Human Reproduction, 2002
    Co-Authors: Hongmei Wang, Yunge Zhao, Qingxiang Amy Sang, Haiyan Lin, Cheng Zhu
    Abstract:

    Matrix Metalloproteinases (MMPs) and their tissue inhibitors may play important roles in tissue remodelling processes of the uterus. This study identified MMP-26 (endometase/matrilysin-2) in the endometrium of pregnant rhesus monkeys (Macaca mulatta) and monitored the spatial and temporal expression of the transcript and protein in the uteri on days 12, 18 and 26 of pregnancy. The partial monkey MMP-26 gene sequence of 289 nucleotides was 98% identical to that of its human homologue and its protein fragment contained a PHCGVPDGSD sequence in the prodomain identical to that in human MMP-26. RT-PCR analysis demonstrated that the average level of MMP-26 mRNA in the endometrium was high on day 12 of pregnancy, but significantly decreased on days 18 and 26 (P < 0.05). In-situ hybridization confirmed that MMP-26 mRNA is specifically localized in the endometrial compartments, with intense signals in the glandular epithelium on day 12 and in the walls of spiral arterioles adjacent to the implantation site on day 26. The hybridization signal for MMP-26 mRNA in the glandular epithelium decreased dramatically on day 18 and was undetectable on day 26. No MMP-26 mRNA transcripts were detected in the placental villi on days 18 and 26. Immunohistochemistry showed that the expression pattern of MMP-26 protein was similar to that of its mRNA. The restricted expression pattern of MMP-26 in the monkey uterus implies that this new MMP is involved in the highly regulated tissue remodelling processes of the glandular epithelium and spiral arteries during early pregnancy.

Alex Y Strongin - One of the best experts on this subject based on the ideXlab platform.

  • Matrix Metalloproteinase 26 proteolysis of the nh2 terminal domain of the estrogen receptor β correlates with the survival of breast cancer patients
    Cancer Research, 2006
    Co-Authors: Alexei Y Savinov, Albert G Remacle, Vladislav S Golubkov, Maryla Krajewska, Susan Kennedy, M J Duffy, Dmitri V Rozanov, Stan Krajewski, Alex Y Strongin
    Abstract:

    Estrogens have many cellular functions, including their interactions with estrogen receptors α and β (ERα and ERβ). Earlier, we determined that the estrogen-ER complex stimulates the transcriptional activity of the Matrix Metalloproteinase 26 ( MMP-26 ) gene promoter. We then determined that ERβ is susceptible to MMP-26 proteolysis whereas ERα is resistant to the protease. MMP-26 targets the NH2-terminal region of ERβ coding for the divergent NH2-terminal A/B domain that is responsible for the ligand-independent transactivation function. As a result, MMP-26 proteolysis generates the COOH-terminal fragments of ERβ. Immunohistochemical analysis of tissue microarrays derived from 121 cancer patients corroborated these data and revealed an inverse correlation between the ERα-dependent expression of MMP-26 and the levels of the intact ERβ in breast carcinomas. MMP-26 is not expressed in normal mammary epithelium. The levels of MMP-26 are strongly up-regulated in ductal carcinoma in situ (DCIS). In the course of further disease progression through stages I to III, the expression of MMP-26 decreases. In contrast to many tumor-promoting MMPs, the expression of MMP-26 in DCIS correlated with a longer patient survival. Our data suggest the existence of an MMP-26–mediated intracellular pathway that targets ERβ and that MMP-26, a novel and valuable cancer marker, contributes favorably to the survival of the ERα/β–positive cohort of breast cancer patients. (Cancer Res 2006; 66(5): 2716-24)

  • Matrix Metalloproteinase 26 is associated with estrogen dependent malignancies and targets α1 antitrypsin serpin
    Cancer Research, 2004
    Co-Authors: Alexei Y Savinov, Vladislav S Golubkov, Dmitri V Rozanov, Hirad Hedayat, Tatiana I Postnova, Natalia V Golubkova, Yu Linli, Stanislaw Krajewski, Alex Y Strongin
    Abstract:

    Proteases exert control over cell behavior and affect many biological processes by making proteolytic modification of regulatory proteins. The purpose of this paper is to describe novel, important functions of Matrix Metalloproteinase (MMP)-26. α1-Antitrypsin (AAT) is a serpin, the primary function of which is to regulate the activity of neutrophil/leukocyte elastase. Insufficient antiprotease activity because of AAT deficiency in the lungs is a contributing factor to early-onset emphysema. We recently discovered that AAT is efficiently cleaved by a novel Metalloproteinase, MMP-26, which exhibits an unconventional PH 81 CGVPD Cys switch motif and is autocatalytically activated in cells and tissues. An elevated expression of MMP-26 in macrophages and polymorphonuclear leukocytes supports the functional role of MMP-26 in the AAT cleavage and inflammation. We have demonstrated a direct functional link of MMP-26 expression with an estrogen dependency and confirmed the presence of the estrogen-response element in the MMP-26 promoter. Immunostaining of tumor cell lines and biopsy specimen microarrays confirmed the existence of the inverse correlations of MMP-26 and AAT in cells/tissues. An expression of MMP-26 in the estrogen-dependent neoplasms is likely to contribute to the inactivation of AAT, to the follow-up liberation of the Ser protease activity, and because of these biochemical events, to promote Matrix destruction and malignant progression. In summary, we hypothesize that MMP-26, by cleaving and inactivating the AAT serpin, operates as a unique functional link that regulates a coordinated interplay between Ser and Metalloproteinases in estrogen-dependent neoplasms.

  • β catenin regulates the gene of mmp 26 a novel Matrix Metalloproteinase expressed both in carcinomas and normal epithelial cells
    The International Journal of Biochemistry & Cell Biology, 2004
    Co-Authors: Natalia D Marchenko, George N Marchenko, Robert N Weinreb, James D Lindsey, Ainura Kyshtoobayeva, Howard C Crawford, Alex Y Strongin
    Abstract:

    Abstract There are several unorthodox features, which distinguish the non-redundant and unique novel Matrix Metalloproteinase-26 (MMP-26) (an enzyme that has recently evolved and does not exist in rodents but is present in humans) from other members of the MMP superfamily. This report describes our recent efforts to gain a better understanding of the mechanisms which restrict expression of MMP-26 to certain cell/tissue types. We examined transcriptional regulation of the human MMP-26 gene in normal and malignant cells. The AP-1 and Tcf-4 sites of the MMP-26 promoter appear most potent in regulating the expression of the MMP-26-luciferase chimera in HEK293 embryonic kidney and MCF7 breast carcinoma cells. Key regulators of the Wnt pathway (β-catenin and lymphoid enhancer-binding factor/T-cell factor with which β-catenin associates) enhanced the transcriptional activity of MMP-26 suggesting that the MMP-26 gene is a likely target of the Wnt pathway. Immunostaining, gene arrays and reverse-transcriptase polymerase chain reaction (RT-PCR) confirm the presence of MMP-26 in normal cells, including the apical epithelial conjunctiva cells of the human eye, as well as in malignant cells of epithelial origin. MMP-26 predominantly accumulates in its proenzyme form in the intracellular milieu of the transfected breast carcinoma MCF7 cells. This study brings us a step forward towards a better understanding of the unconventional role, regulation and functions of epithelial cell MMP-26 in physiological conditions and in neoplasms.

  • promoter characterization of the novel human Matrix Metalloproteinase 26 gene regulation by the t cell factor 4 implies specific expression of the gene in cancer cells of epithelial origin
    Biochemical Journal, 2002
    Co-Authors: George N Marchenko, Natalia D Marchenko, Jay Leng, Alex Y Strongin
    Abstract:

    A novel Matrix Metalloproteinase-26 (MMP-26) is known to be specifically expressed in epithelial carcinomas. To facilitate studies of MMP-26 transcriptional regulation, we have cloned and characterized a 1 kb 5'-flanking region of the human MMP-26 gene. Altogether, our findings indicate that the MMP-26 promoter has distinctive structural and functional features among MMP genes. An unusual polyadenylation site proximal to the transcription-factor-binding sites protects transcription of the MMP-26 gene from the upstream promoters and represents a part of the stringent transcriptional regulation of the gene. The MMP-26 gene has a consensus TATA-box and one transcriptional start site located 60 and 35 nucleotides upstream of the translational start site, respectively. The MMP-26 promoter was able to drive luciferase expression in human A549 lung carcinoma, HT1080 fibrosarcoma and HEK293 embryonic kidney cells. The basal transcription efficiency of the MMP-26 promoter is relatively low, thereby explaining the minute expression of the gene in most cells and tissues. When compared with other MMP genes, the MMP-26 promoter contains binding sites for a few transcription factors. Sequential deletion and mutation analysis, and electrophoretic mobility-shift assay have identified the T-cell factor-4 (Tcf-4) motif and the activator protein-1 site as the major regulatory elements of the MMP-26 promoter. Since previous studies have established that the Tcf-4 transcription factor is subjected exclusively to regulation through the beta-catenin/E(epithelial)-cadherin pathway, this implies the specific expression of MMP-26 in cancer cells of epithelial origin.

  • characterization of Matrix Metalloproteinase 26 a novel Metalloproteinase widely expressed in cancer cells of epithelial origin
    Biochemical Journal, 2001
    Co-Authors: George N Marchenko, Boris I Ratnikov, Dmitry V Rozanov, Adam Godzik, Elena I Deryugina, Alex Y Strongin
    Abstract:

    Identification of expanding roles for Matrix Metalloproteinases (MMPs) in complex regulatory processes of tissue remodelling has stimulated the search for genes encoding proteinases with unique functions, regulation and expression patterns. By using a novel cloning strategy, we identified three previously unknown human MMPs, i.e. MMP-21, MMP-26 and MMP-28, in comprehensive gene libraries. The present study is focused on the gene and the protein of a novel MMP, MMP-26. Our findings show that MMP-26 is specifically expressed in cancer cells of epithelial origin, including carcinomas of lung, prostate and breast. Several unique structural and regulatory features, including an unusual 'cysteine-switch' motif, discriminate broad-spectrum MMP-26 from most other MMPs. MMP-26 efficiently cleaves fibrinogen and extracellular Matrix proteins, including fibronectin, vitronectin and denatured collagen. Protein sequence, minimal modular domain structure, exon-intron mapping and computer modelling demonstrate similarity between MMP-26 and MMP-7 (matrilysin). However, substrate specificity and transcriptional regulation, as well as the functional role of MMP-26 and MMP-7 in cancer, are likely to be distinct. Despite these differences, matrilysin-2 may be a suitable trivial name for MMP-26. Our observations suggest an important specific function for MMP-26 in tumour progression and angiogenesis, and confirm and extend the recent findings of other authors [Park, Ni, Gerkema, Liu, Belozerov and Sang (2000) J. Biol. Chem. 275, 20540--20544; Uria and Lopez-Otin (2000) Cancer Res. 60, 4745--4751; de Coignac, Elson, Delneste, Magistrelli, Jeannin, Aubry, Berthier, Schmitt, Bonnefoy and Gauchat (2000) Eur. J. Biochem. 267, 3323--3329].

Hyun I Park - One of the best experts on this subject based on the ideXlab platform.

  • the intermediate s1 pocket of the endometase matrilysin 2 active site revealed by enzyme inhibition kinetic studies protein sequence analyses and homology modeling
    Journal of Biological Chemistry, 2003
    Co-Authors: Hyun I Park, Seakwoo Lee, Douglas R. Hurst, Yonghao Jin, Cyrus A Monroe, Martin A Schwartz, Qingxiang Amy Sang
    Abstract:

    Human Matrix Metalloproteinase-26 (MMP-26/endometase/matrilysin-2) is a newly identified MMP and its structure has not been reported. The enzyme active site S1′ pocket in MMPs is a well defined substrate P1′ amino acid residue-binding site with variable depth. To explore MMP-26 active site structure-activity, a series of new potent mercaptosulfide MMP inhibitors (MMPIs) with Leu or homophenylalanine (Homophe) side chains at the P1′ site were selected. The Homephe side chain is designed to probe deep S1′ pocket MMPs. These inhibitors were tested against MMP-26 and several MMPs with known x-ray crystal structures to distinguish shallow, intermediate, and deep S1′ pocket characteristics. MMP-26 has an inhibition profile most similar to those of MMPs with intermediate S1′ pockets. Investigations with hydroxamate MMPIs, including those designed for deep pocket MMPs, also indicated the presence of an intermediate pocket. Protein sequence analysis and homology modeling further verified that MMP-26 has an intermediate S1′ pocket formed by Leu-204, His-208, and Tyr-230. Moreover, residue 233 may influence the depth of an MMP S1′ pocket. The residue at the equivalent position of MMP-26 residue 233 is hydrophilic in intermediate-pocket MMPs (e.g. MMP-2, -8, and -9) and hydrophobic in deep-pocket MMPs (e.g. MMP-3, -12, and -14). MMP-26 contains a His-233 that renders the S1′ pocket to an intermediate size. This study suggests that MMPIs, protein sequence analyses, and molecular modeling are useful tools to understand structure-activity relationships and provides new insight for rational inhibitor design that may distinguish MMPs with deep versus intermediate S1′ pockets.

  • activation of pro gelatinase b by endometase matrilysin 2 promotes invasion of human prostate cancer cells
    Journal of Biological Chemistry, 2003
    Co-Authors: Yunge Zhao, Hyun I Park, Aizhen Xiao, Robert G Newcomer, Tiebang Kang, Leland W K Chung, Mark G Swanson, Haiyen E Zhau, John Kurhanewicz, Qingxiang Amy Sang
    Abstract:

    Abstract This work has explored a putative biochemical mechanism by which endometase/matrilysin-2/Matrix Metalloproteinase-26 (MMP-26) may promote human prostate cancer cell invasion. Here, we showed that the levels of MMP-26 protein in human prostate carcinomas from multiple patients were significantly higher than those in prostatitis, benign prostate hyperplasia, and normal prostate glandular tissues. The role of MMP-26 in prostate cancer progression is unknown. MMP-26 was capable of activating pro-MMP-9 by cleavage at the Ala93–Met94 site of the prepro-enzyme. This activation proceeded in a time- and dose-dependent manner, facilitating the efficient cleavage of fibronectin by MMP-9. The activated MMP-9 products generated by MMP-26 appeared more stable than those cleaved by MMP-7 under the conditions tested. To investigate the contribution of MMP-26 to cancer cell invasion via the activation of MMP-9, highly invasive and metastatic human prostate carcinoma cells, androgen-repressed prostate cancer (ARCaP) cells were selected as a working model. ARCaP cells express both MMP-26 and MMP-9. Specific anti-MMP-26 and anti-MMP-9 functional blocking antibodies both reduced the invasiveness of ARCaP cells across fibronectin or type IV collagen. Furthermore, the introduction of MMP-26antisense cDNA into ARCaP cells significantly reduced the MMP-26 protein level in these cells and strongly suppressed the invasiveness of ARCaP cells. Double immunofluorescence staining and confocal laser scanning microscopic images revealed that MMP-26 and MMP-9 were co-localized in parental and MMP-26 sense-transfected ARCaP cells. Moreover, MMP-26 and MMP-9 proteins were both expressed in the same human prostate carcinoma tissue samples examined. These results indicate that MMP-26 may be a physiological and pathological activator of pro-MMP-9.

  • peptide substrate specificities and protein cleavage sites of human endometase matrilysin 2 Matrix Metalloproteinase 26
    Journal of Biological Chemistry, 2002
    Co-Authors: Hyun I Park, Ferry E Gerkema, Benjamin E Turk, Lewis C Cantley, Qingxiang Amy Sang
    Abstract:

    Human endometase/matrilysin-2/Matrix Metalloproteinase-26 (MMP-26) is a novel epithelial and cancer-specific Metalloproteinase. Peptide libraries were used to profile the substrate specificity of MMP-26 from the P4-P4' sites. The optimal cleavage motifs for MMP-26 were Lys-Pro-Ile/Leu-Ser(P1)-Leu/Met(P1')-Ile/Thr-Ser/Ala-Ser. The strongest preference was observed at the P1' and P2 sites where hydrophobic residues were favored. Proline was preferred at P3, and Serine was preferred at P1. The overall specificity was similar to that of other MMPs with the exception that more flexibility was observed at P1, P2', and P3'. Accordingly, synthetic inhibitors of gelatinases and collagenases inhibited MMP-26 with similar efficacy. A pair of stereoisomers had only a 40-fold difference in K(i)(app) values against MMP-26 compared with a 250-fold difference against neutrophil collagenase, indicating that MMP-26 is less stereoselective for its inhibitors. MMP-26 autodigested itself during the folding process. Two of the major autolytic sites were Leu(49)-Thr(50) and Ala(75)-Leu(76), which still left the cysteine switch sequence (PHC(82)GVPD) intact. This suggests that Cys(82) may not play a role in the latency of the zymogen. Interestingly, inhibitor titration studies revealed that only approximately 5% of the total MMP-26 molecules was catalytically active, indicating that the thiol groups of Cys(82) in the active molecules may be dissociated or removed from the active site zinc ions. MMP-26 cleaved Phe(352)-Leu(353) and Pro(357)-Met(358) in the reactive loop of alpha(1)-proteinase inhibitor and His(140)-Val(141) in insulin-like growth factor-binding protein-1, probably rendering these substrates inactive. Among the fluorescent peptide substrates analyzed, Mca-Pro-Leu-Ala-Nva-Dpa-Ala-Arg-NH(2) displayed the highest specificity constant (30,000/molar second) with MMP-26. This report proposes a working model for the future studies of pro-MMP-26 activation, the design of inhibitors, and the identification of optimal physiological and pathological substrates of MMP-26 in vivo.

  • identification and characterization of human endometase Matrix Metalloproteinase 26 from endometrial tumor
    Journal of Biological Chemistry, 2000
    Co-Authors: Hyun I Park, Ferry E Gerkema, Ding Liu, Vladimir E Belozerov, Qingxiang Amy Sang
    Abstract:

    Abstract We report the discovery, cloning, and characterization of a novel human Matrix Metalloproteinase 26 (MMP-26) (Matrixin) gene, endometase, an endometrial tumor-derivedMetalloproteinase. Among more than three million expressed sequence tags sequenced, the endometase gene was only obtained from human endometrial tumor cDNA library. Endometase mRNA was expressed specifically in human uterus, not in other tissues/cells tested, e.g. testis, heart, brain, lungs, liver, thymus, and melanoma G361. Endometase protein has a signal peptide, a propeptide domain, and a catalytic domain with a unique “cysteine switch” propeptide sequence, PHCGVPDGSD, and a zinc-binding motif, VATHEIGHSLGLQH. Endometase is 43, 41, 41, and 39% identical to human metalloelastase, stromelysin, collagenase-3, and matrilysin, respectively. The zymogen was expressed and isolated from Escherichia coli as inclusion bodies with a molecular mass of 28 kDa. The identity and homogeneity of the recombinant protein was confirmed by protein N-terminal sequencing, silver stain, and immunoblot analyses. The pro-enzyme was partially activated during the folding process. Endometase selectively cleaved type I gelatin and α1-proteinase inhibitor; however, it did not digest collagens, laminin, elastin, β-casein, plasminogen, soybean trypsin inhibitor, or Bowman-Birk inhibitor. It hydrolyzed peptide substrates of Matrixins and tumor necrosis factor-α converting enzyme. Endometase may selectively cleave extracellular Matrix proteins, inactivate serpins, and process cytokines.

Keiichi Isaka - One of the best experts on this subject based on the ideXlab platform.

  • estrogen and estrogen receptor induce Matrix Metalloproteinase 26 expression in endometrial carcinoma cells
    Oncology Reports, 2013
    Co-Authors: Hirotaka Nishi, Masahiko Kuroda, Keiichi Isaka
    Abstract:

    The human Matrix Metalloproteinase (MMP)-26, also called matrilysin-2 or endometase, has been isolated as a matrilysin (MMP-7) homolog. Several reports describe that MMP-26 may be related to the development of endometrial carcinomas. Total RNAs were isolated from 51 normal endometrial tissue samples, 6 endometrial hyperplasia tissue samples and 30 endometrial carcinomas. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate MMP-26 mRNA expression levels. We examined the effect of estrogen and its receptor (ER) on MMP-26 expression in endometrial carcinoma cell lines by real-time RT-PCR, western blot analysis and luciferase assays. To examine protein-DNA binding between ER and MMP-26 promoter, we performed chromatin immunoprecipitation (ChIP) assay. Real-time RT-PCR analysis revealed that MMP-26 mRNA expression was significantly higher in the normal human endometria and hyperplasias compared with that in endometrial carcinomas. Estrogen not only transactivated the MMP-26 promoter activity but also enhanced endogenous MMP-26 expression. The MMP-26 promoter region contains a putative ER response element (ERE). Nuclear ER protein interacted with ERE on the MMP-26 promoter by ChIP assay. We found a significant difference in MMP-26 expression in normal and malignant endometrial tissue samples and that estrogen induced MMP-26 expression. Estrogen may induce endometrial hyperplasia but not endometrial carcinoma. Our results provide evidence that regulation of MMP-26 promoter activity by estrogen may represent a mechanism for endometrial carcinogenesis.

  • Matrix Metalloproteinase 26 is present more frequently in squamous cell carcinomas of immunosuppressed compared with immunocompetent patients
    Journal of Cutaneous Pathology, 2009
    Co-Authors: Tiina Kuivanen, Keiichi Isaka, Leila Jeskanen, L Kyllonen, Ulpu Saarialhokere
    Abstract:

    Background: Skin cancers are the most frequent malignancies in organ transplant recipients (OTRs). Squamous cell carcinomas (SCCs) occur 65–250 times more frequently in OTRs and tend to be aggressive in behavior. Because Matrix Metalloproteinases (MMPs) have a central role in tumorigenesis and invasion, we investigated the epithelial and stromal MMP and tissue inhibitor of MMP (TIMP) expression profile in SCCs of immunosuppressed (IS) compared with immunocompetent (IC) patients to determine if differences could explain the more aggressive behavior of SCCs in OTRs. Methods: Matched pairs from 20 SCCs of IS and IC patients were studied using immunohistochemistry for MMP-1, MMP-7, MMP-8, MMP-9, MMP-13 and MMP-26 and TIMP-1 and TIMP-3. Results: Among all MMPs studied, only staining for MMP-26 was significantly more intense in cancer cells of the post-transplant group compared with the IC group (p = 0.01), whereas MMP-9 expression was more abundant in stromal macrophages surrounding SCCs of IC patients (p = 0.02). MMP-26 expression in cancer cells (p = 0.04) and that of MMP-9 in neutrophils (p = 0.005) were more abundant in SCCs of patients using cyclosporine. Conclusions: We conclude that MMP-26 and MMP-9 may contribute to the more aggressive behavior of SCCs in OTRs.

  • matrilysin 2 Matrix Metalloproteinase 26 is upregulated in keratinocytes during wound repair and early skin carcinogenesis
    Journal of Investigative Dermatology, 2005
    Co-Authors: Katja Ahokas, Tiina Skoog, Sari Suomela, Leila Jeskanen, Ulla Impola, Keiichi Isaka
    Abstract:

    Matrilysin-2 (Matrix Metalloproteinase (MMP)-26) is a small protein of the MMP family expressed in some epithelial carcinomas and normal tissues. We studied its role in benign skin disorders characterized by epithelial proliferation, in wound repair, skin cancer, and regulation in keratinocyte (KC) cultures. MMP-26 is expressed by laminin-5-positive KC in the migrating area during wound repair, in benign skin disorders characterized by inflammation and microdisruptions of basement membrane, but in intact skin only in hair follicles. It was detected in occasional atypical KC in pre-malignant lesions but not in basal cell cancer islands. Although MMP-26 was expressed in grades I and II squamous cell cancers (SCC), it was not present in dedifferentiated grade III tumors. MMP-26 was neither co-expressed with its close homologue matrilysin-1 nor with the proliferation marker Ki-67. But in tissue samples it either co-localized or was detected in adjacent cells of same regions with the tumor suppressor p16. In KC and HaCaT cell cultures, 12-phorbol-13-myristate-acetate, epidermal growth factor, tumor necrosis factor-α, transforming growth factor-β1, interleukin-1 (IL-1)β, IL-6, insulin-like growth factor, γ-IFN, retinoic acid, dexamethasone, four matrices or ras -transformation were unable to upregulate MMP-26 expression. The expression pattern of MMP-26 suggests that it may be upregulated in basal KC even without tumorigenesis because of altered cell–Matrix interactions and inflammation and, unlike most MMP, becomes downregulated during histological dedifferentiation of SCC. Thus, lack of MMP-26 in SCC could be a marker of aggressive growth.

  • Matrix Metalloproteinase 26 is expressed in human endometrium but not in endometrial carcinoma
    Cancer, 2003
    Co-Authors: Keiichi Isaka, Hirotaka Nishi, M Hiromi D Nakai, M Toshihide D Nakada, Yoshiro Ebihara, Masaomi Takayama
    Abstract:

    BACKGROUND The human Matrix Metalloproteinase (MMP)-26, also called matrilysin-2 or endometase, has been isolated as a matrilysin (MMP-7) homolog. Matrix Metalloproteinase-26 was expressed in tissue samples from the placenta and endometrial tumors and its expression may be related to the development of endometrial carcinomas. METHODS Total RNAs were isolated from 5 endometrial carcinoma cell lines, 36 normal endometrial tissue samples, 4 hyperplasia tissue samples, and from 24 endometrial carcinoma tissue samples. Reverse transcription-polymerase chain reation (RT-PCR) was performed to detect MMP-26 mRNA expression. To identify MMP-26 mRNA localization and protein expression, we performed in situ RT-PCR and immunohistochemistry, respectively. RESULTS Reverse transcription-polymerase chain reaction analysis revealed that MMP-26 mRNA was expressed in 24 of 36 normal human endometrial tissue samples. However, MMP-26 mRNA expression was not detected in endometrial carcinoma cell lines nor in endometrial carcinoma tissue samples except for one case. Western blot analysis showed similar results. In situ RT-PCR analysis revealed that MMP-26 expression was localized in the epithelial glandular cells but faint expression was observed in the stromal cells. Subsequently, we separated endometrial tissues into epithelial glandular and stromal cells. Using RT-PCR, the purified epithelial glandular cells exhibited MMP-26 mRNA expression but the purified stromal cells did not. Immunohistochemical analyses revealed that MMP-26 protein expression is also limited to endometrial epithelial glandular cells but not to cancer cells. Therefore, MMP-26 expression is limited to normal epithelial glandular cells. CONCLUSIONS We found a significant difference in MMP-26 expression in normal and malignant endometrial tissue samples, although its function is still unknown. These data suggest that MMP-26 may be a candidate for a new tumor marker for endometrial carcinomas. Cancer 2003;97:79–89. © 2003 American Cancer Society. DOI 10.1002/cncr.11030