The Experts below are selected from a list of 6531 Experts worldwide ranked by ideXlab platform
Osamu Yoshida - One of the best experts on this subject based on the ideXlab platform.
-
single step purification of Prostatic Acid Phosphatase immunoaffinity chromatography with a monoclonal antibody
International Journal of Urology, 1995Co-Authors: Tatsuhiro Yoshiki, Masamichi Ueda, Atsushi Hirano, Kenichiro Okada, Osamu YoshidaAbstract:Background: Prostatic Acid Phosphatase (PAP) is an important protein which should be studied further as a tumor marker or as a biologically functional molecule. The purpose of the study was to establish a simple and reliable method to obtain highly pure PAP. Methods: Spleen cells from mice immunized with Prostatic epithelial cells prepared from benign Prostatic hyperplasia tissue were fused with myeloma cells X63Ag8–653. Hybrid cells of interest were selected using the indirect immunofluorescence method with unfixed frozen tissue sections. One clone of the hybrid cell lines was established which secreted the monoclonal antibody specifically reactive to Prostatic Acid Phosphatase. Using this monoclonal antibody, we purified the antigen from human Prostatic tissue by means of single-step immunoaffinity chromatography. Results: SDS-PAGE profiling under reducing conditions indicated that the protein recognized by this antibody consisted of several components of molecular weight 41,000–45,000. Partial amino Acid sequence analysis of this protein indicated that these components involved a heterogeneously modified single polypeptide, and that this antigen is identical to human Prostatic Acid Phosphatase. Conclusions: This single-step method saves the time needed to purify Prostatic Acid Phosphatase and requires only half a day for the whole procedure. Moreover, the purity of the isolated protein was extremely high. This method seems to be useful not only for purifying Prostatic Acid Phosphatase but also for purifying other proteins from the prostate gland and for analysis of antigenic macromolecules.
-
SINGLE‐STEP PURIFICATION OF Prostatic Acid Phosphatase: IMMUNOAFFINITY CHROMATOGRAPHY WITH A MONOCLONAL ANTIBODY
International Journal of Urology, 1995Co-Authors: Tatsuhiro Yoshiki, Masamichi Ueda, Atsushi Hirano, Kenichiro Okada, Osamu YoshidaAbstract:Background: Prostatic Acid Phosphatase (PAP) is an important protein which should be studied further as a tumor marker or as a biologically functional molecule. The purpose of the study was to establish a simple and reliable method to obtain highly pure PAP. Methods: Spleen cells from mice immunized with Prostatic epithelial cells prepared from benign Prostatic hyperplasia tissue were fused with myeloma cells X63Ag8–653. Hybrid cells of interest were selected using the indirect immunofluorescence method with unfixed frozen tissue sections. One clone of the hybrid cell lines was established which secreted the monoclonal antibody specifically reactive to Prostatic Acid Phosphatase. Using this monoclonal antibody, we purified the antigen from human Prostatic tissue by means of single-step immunoaffinity chromatography. Results: SDS-PAGE profiling under reducing conditions indicated that the protein recognized by this antibody consisted of several components of molecular weight 41,000–45,000. Partial amino Acid sequence analysis of this protein indicated that these components involved a heterogeneously modified single polypeptide, and that this antigen is identical to human Prostatic Acid Phosphatase. Conclusions: This single-step method saves the time needed to purify Prostatic Acid Phosphatase and requires only half a day for the whole procedure. Moreover, the purity of the isolated protein was extremely high. This method seems to be useful not only for purifying Prostatic Acid Phosphatase but also for purifying other proteins from the prostate gland and for analysis of antigenic macromolecules.
Hiroyuki Arai - One of the best experts on this subject based on the ideXlab platform.
-
Prostatic Acid Phosphatase degrades lysophosphatidic Acid in seminal plasma
FEBS Letters, 2004Co-Authors: Masayuki Tanaka, Yasuhiro Kishi, Yasukazu Takanezawa, Yoshiyuki Kakehi, Junken Aoki, Hiroyuki AraiAbstract:Lysophosphatidic Acid (LPA) is a lipid mediator with multiple biological activities and is detected in various biological fluids, including human seminal plasma. Due to its cell proliferation stimulatory and anti-apoptotic activities, LPA has been implicated in the progression of some cancers such as ovarian cancer and prostate cancer. Here, we show that Prostatic Acid Phosphatase, which is a non-specific Phosphatase and which has been implicated in the progression of prostate cancer, inactivates LPA in human seminal plasma. Human seminal plasma contains both an LPA-synthetic enzyme, lysoPLD, which converts lysophospholipids to LPA and is responsible for LPA production in serum, and its major substrate, lysophosphatidylcholine. In serum, LPA accumulated during incubation at 37 °C. However, in seminal plasma, LPA did not accumulate. This discrepancy is explained by the presence of a strong LPA-degrading activity. Incubation of LPA with seminal plasma resulted in the disappearance of LPA and an accompanying accumulation of monoglyceride showing that LPA is degraded by Phosphatase activity present in the seminal plasma. When seminal plasma was incubated in the presence of a Phosphatase inhibitor, sodium orthovanadate, LPA accumulated, indicating that LPA is produced and degraded in the fluid. Biochemical characterization of the LPA-Phosphatase activity identified two Phosphatase activities in human seminal plasma. By Western blotting analysis in combination with several column chromatographies, the major activity was revealed to be identical to Prostatic Acid Phosphatase. The present study demonstrates active LPA metabolism in seminal plasma and indicates the possible role of LPA signaling in male sexual organs including prostate cancer.
Lukasz Lebioda - One of the best experts on this subject based on the ideXlab platform.
-
Structural Origins of l(+)-Tartrate Inhibition of Human Prostatic Acid Phosphatase
Journal of Biological Chemistry, 1998Co-Authors: Michael W. Lacount, George Handy, Lukasz LebiodaAbstract:Acid Phosphatase activity in the blood serum is usually separated into tartrate-resistant and tartrate-refractory, which is reported as the Prostatic Acid Phosphatase level. Human Prostatic Acid Phosphatase crystals soaked in N-propyl-L-tartramate were used to collect x-ray diffraction data to 2.9 A resolution under cryogenic conditions. Positive difference electron density, corresponding to the inhibitor, was found. The quality of the electron density maps clearly shows the orientation of the carboxylate and N-propyl-substituted amide groups. The hydroxyl group attached to C3 forms two crucial hydrogen bonds with Arg-79 and His-257. Previous crystallographic studies compiled on the tartrate-rat Prostatic Acid Phosphatase binary complex (Lindqvist, Y., Schneider, G., and Vihko, P. (1993) J. Biol. Chem. 268, 20744-20746) erroneously positioned D-tartrate into the active site. Modeling studies have shown that the C3 hydroxyl group on the D(-)-stereoisomer of tartrate, which does not significantly inhibit Prostatic Acid Phosphatase, does not form strong hydrogen bonds with Arg-79 or His-257. The structure of human Prostatic Acid Phosphatase, noncovalently bound in N-propyl-L-tartramate, is used to develop inhibitors with higher specificity and potency than L(+)-tartrate.
-
structural origins of l tartrate inhibition of human Prostatic Acid Phosphatase
Journal of Biological Chemistry, 1998Co-Authors: Michael W. Lacount, George Handy, Lukasz LebiodaAbstract:Acid Phosphatase activity in the blood serum is usually separated into tartrate-resistant and tartrate-refractory, which is reported as the Prostatic Acid Phosphatase level. Human Prostatic Acid Phosphatase crystals soaked in N-propyl-L-tartramate were used to collect x-ray diffraction data to 2.9 A resolution under cryogenic conditions. Positive difference electron density, corresponding to the inhibitor, was found. The quality of the electron density maps clearly shows the orientation of the carboxylate and N-propyl-substituted amide groups. The hydroxyl group attached to C3 forms two crucial hydrogen bonds with Arg-79 and His-257. Previous crystallographic studies compiled on the tartrate-rat Prostatic Acid Phosphatase binary complex (Lindqvist, Y., Schneider, G., and Vihko, P. (1993) J. Biol. Chem. 268, 20744-20746) erroneously positioned D-tartrate into the active site. Modeling studies have shown that the C3 hydroxyl group on the D(-)-stereoisomer of tartrate, which does not significantly inhibit Prostatic Acid Phosphatase, does not form strong hydrogen bonds with Arg-79 or His-257. The structure of human Prostatic Acid Phosphatase, noncovalently bound in N-propyl-L-tartramate, is used to develop inhibitors with higher specificity and potency than L(+)-tartrate.
Robert A Gardiner - One of the best experts on this subject based on the ideXlab platform.
-
tartrate inhibition of Prostatic Acid Phosphatase improves seminal fluid metabolite stability
Metabolomics, 2016Co-Authors: Matthew J Roberts, Jake P N Hattwell, Clement W K Chow, Martin F Lavin, Gregory K Pierens, Robert A Gardiner, Horst Joachim SchirraAbstract:Introduction Human seminal fluid (hSF) has been suggested as a biofluid suitable to characterise male reproductive organ pathology with metabolomics. However, various enzymatic processes, including phosphorylcholine hydrolysis mediated by Prostatic Acid Phosphatase (PAP), cause unwanted metabolite variation that may complicate metabolomic analysis of fresh hSF samples.
Matthew J Roberts - One of the best experts on this subject based on the ideXlab platform.
-
tartrate inhibition of Prostatic Acid Phosphatase improves seminal fluid metabolite stability
Metabolomics, 2016Co-Authors: Matthew J Roberts, Jake P N Hattwell, Clement W K Chow, Martin F Lavin, Gregory K Pierens, Robert A Gardiner, Horst Joachim SchirraAbstract:Introduction Human seminal fluid (hSF) has been suggested as a biofluid suitable to characterise male reproductive organ pathology with metabolomics. However, various enzymatic processes, including phosphorylcholine hydrolysis mediated by Prostatic Acid Phosphatase (PAP), cause unwanted metabolite variation that may complicate metabolomic analysis of fresh hSF samples.
