The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
M. S. Weiss - One of the best experts on this subject based on the ideXlab platform.
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structure based epitope mapping of Mycobacterium Tuberculosis secretary antigen mtc28
Journal of Biological Chemistry, 2016Co-Authors: Prasun Kundu, Rupam Biswas, Linda Reinhard, Anirudha Dutta, Jochen Muellerdieckmann, Somnath Mukherjee, M. S. WeissAbstract:Abstract Secretary proteins of Mycobacterium Tuberculosis are the key players of mycobacterial infection pathway. MTC28 is a 28kDa proline-rich secretary antigen of Mycobacterium Tuberculosis and is only conserved in pathogenic strains of mycobacteria. Here we report the crystal structure of MTC28 at 2.8 and 2.15 angstrom resolutions for structure based epitope design. MTC28 shares a mog1p fold consisting of seven antiparallel beta strands stacked between alpha helices. Five probable epitopes have been located on a solvent accessible flexible region by computational analysis of the structure of MTC28. Simultaneously, the protein is digested with trypsin and the resulting fragments are purified by HPLC. Such 10 purified peptide fragments are screened against sera from patients infected with pulmonary Tuberculosis (PTB). Two out of these ten fragments namely128ALDITLPMPPR137 and 138WTQVPDPNVPDAFVVIADR157 are found to be major immunogenic epitopes that are localized on the outer surface of the protein molecule and are part of a single continuous epitope that have been predicted in silico. Mutagenesis and antibody inhibition studies are in accordance with the results obtained from epitope mapping.
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structure based epitope mapping of Mycobacterium Tuberculosis secretary antigen mtc28
Journal of Biological Chemistry, 2016Co-Authors: Prasun Kundu, Rupam Biswas, Linda Reinhard, Anirudha Dutta, Jochen Muellerdieckmann, M. S. Weiss, Somnath Mukherjee, Nishith Kumar Pal, Amit DasAbstract:Secretary proteins of Mycobacterium Tuberculosis are key players of the mycobacterial infection pathway. MTC28 is a 28-kDa proline-rich secretary antigen of Mycobacterium Tuberculosis and is only conserved in pathogenic strains of mycobacteria. Here we report the crystal structure of MTC28 at 2.8- and 2.15-A resolutions for the structure-based epitope design. MTC28 shares a "mog1p"-fold consisting of seven antiparallel β strands stacked between α helices. Five probable epitopes have been located on a solvent-accessible flexible region by computational analysis of the structure of MTC28. Simultaneously, the protein is digested with trypsin and the resulting fragments are purified by HPLC. Such 10 purified peptide fragments are screened against sera from patients infected with pulmonary Tuberculosis (PTB). Two of these 10 fragments, namely (127)ALDITLPMPPR(137) and (138)WTQVPDPNVPDAFVVIADR(156),are found to be major immunogenic epitopes that are localized on the outer surface of the protein molecule and are part of a single continuous epitope that have been predicted in silico Mutagenesis and antibody inhibition studies are in accordance with the results obtained from epitope mapping.
Prasun Kundu - One of the best experts on this subject based on the ideXlab platform.
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structure based epitope mapping of Mycobacterium Tuberculosis secretary antigen mtc28
Journal of Biological Chemistry, 2016Co-Authors: Prasun Kundu, Rupam Biswas, Linda Reinhard, Anirudha Dutta, Jochen Muellerdieckmann, Somnath Mukherjee, M. S. WeissAbstract:Abstract Secretary proteins of Mycobacterium Tuberculosis are the key players of mycobacterial infection pathway. MTC28 is a 28kDa proline-rich secretary antigen of Mycobacterium Tuberculosis and is only conserved in pathogenic strains of mycobacteria. Here we report the crystal structure of MTC28 at 2.8 and 2.15 angstrom resolutions for structure based epitope design. MTC28 shares a mog1p fold consisting of seven antiparallel beta strands stacked between alpha helices. Five probable epitopes have been located on a solvent accessible flexible region by computational analysis of the structure of MTC28. Simultaneously, the protein is digested with trypsin and the resulting fragments are purified by HPLC. Such 10 purified peptide fragments are screened against sera from patients infected with pulmonary Tuberculosis (PTB). Two out of these ten fragments namely128ALDITLPMPPR137 and 138WTQVPDPNVPDAFVVIADR157 are found to be major immunogenic epitopes that are localized on the outer surface of the protein molecule and are part of a single continuous epitope that have been predicted in silico. Mutagenesis and antibody inhibition studies are in accordance with the results obtained from epitope mapping.
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structure based epitope mapping of Mycobacterium Tuberculosis secretary antigen mtc28
Journal of Biological Chemistry, 2016Co-Authors: Prasun Kundu, Rupam Biswas, Linda Reinhard, Anirudha Dutta, Jochen Muellerdieckmann, M. S. Weiss, Somnath Mukherjee, Nishith Kumar Pal, Amit DasAbstract:Secretary proteins of Mycobacterium Tuberculosis are key players of the mycobacterial infection pathway. MTC28 is a 28-kDa proline-rich secretary antigen of Mycobacterium Tuberculosis and is only conserved in pathogenic strains of mycobacteria. Here we report the crystal structure of MTC28 at 2.8- and 2.15-A resolutions for the structure-based epitope design. MTC28 shares a "mog1p"-fold consisting of seven antiparallel β strands stacked between α helices. Five probable epitopes have been located on a solvent-accessible flexible region by computational analysis of the structure of MTC28. Simultaneously, the protein is digested with trypsin and the resulting fragments are purified by HPLC. Such 10 purified peptide fragments are screened against sera from patients infected with pulmonary Tuberculosis (PTB). Two of these 10 fragments, namely (127)ALDITLPMPPR(137) and (138)WTQVPDPNVPDAFVVIADR(156),are found to be major immunogenic epitopes that are localized on the outer surface of the protein molecule and are part of a single continuous epitope that have been predicted in silico Mutagenesis and antibody inhibition studies are in accordance with the results obtained from epitope mapping.
T J Cleary - One of the best experts on this subject based on the ideXlab platform.
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evaluation of gen probe amplified Mycobacterium Tuberculosis direct test and pcr for direct detection of Mycobacterium Tuberculosis in clinical specimens
Journal of Clinical Microbiology, 1994Co-Authors: Nancimae Miller, S G Hernandez, T J ClearyAbstract:The Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (AMTD) is a direct specimen assay for the identification of Mycobacterium Tuberculosis from respiratory samples. rRNA is amplified, and the product is detected with a specific chemiluminescent probe. We performed a retrospective evaluation of three separate respiratory specimens from each of 250 patients by using the AMTD and compared the results with those of microscopy, culturing, and a patient chart review. From the latter results, 198 patients (594 specimens) were found negative for M. Tuberculosis by culturing and clinical criteria. The overall specificity of the AMTD after discrepancy resolution was 98.5% (585 of 594). There were 52 patients with culture-proven and/or clinically diagnosed Tuberculosis. Of these 156 specimens, the organism was cultured from 142 (91%), and acid-fast microscopy was positive for 105 (67.3%). The AMTD was positive for 142 (91%) specimens from these patients. Tuberculosis patient samples were tested by a PCR assay which uses primers for amplification of the IS6110 insertion sequence of the M. Tuberculosis complex. The PCR assay detected 144 of the 156 (92.3%) specimens. Overall, when three specimens per patient were examined, the AMTD found all 52 patients positive for Tuberculosis, while the PCR assay found 51 patients positive by agarose gel analysis and all 52 patients positive by Southern blot hybridization.
S Goto - One of the best experts on this subject based on the ideXlab platform.
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detection of Mycobacterium Tuberculosis in clinical specimens by polymerase chain reaction and gen probe amplified Mycobacterium Tuberculosis direct test
Journal of Clinical Microbiology, 1993Co-Authors: Chiyoji Abe, Kazue Hirano, Masako Wada, Y Kazumi, Mitsuyoshi Takahashi, Y Fukasawa, T Yoshimura, C Miyagi, S GotoAbstract:The polymerase chain reaction (PCR) using oligonucleotides based on the repetitive sequence (IS986) of Mycobacterium Tuberculosis as a primer and the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD), which combines an M. Tuberculosis rRNA amplification method with the hybridization protection assay format, were evaluated for detection of M. Tuberculosis in clinical samples. The detection limits of these two assay systems based on nucleic acid amplification for cultured M. Tuberculosis were less than 10 cells per reaction. A total of 135 sputum specimens were examined by the two assay systems. The PCR and the MTD systems for detection of M. Tuberculosis gave overall positivity rates of 84.2% (32 of 38) and 91.9% (34 of 37), respectively, as compared with 71.9% (23 of 32) by smear and 96.9% (31 of 32) by culture in the liquid medium MB-Check. Procedures for sample preparation used in the two methods were different. Although the sensitivities of the PCR and MTD appeared to be similar to that of culture with the MB-Check system, the two methods based on nucleic acid amplification should be very useful for rapid detection of M. Tuberculosis infections without the long time required for culture of M. Tuberculosis.
Amit Das - One of the best experts on this subject based on the ideXlab platform.
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structure based epitope mapping of Mycobacterium Tuberculosis secretary antigen mtc28
Journal of Biological Chemistry, 2016Co-Authors: Prasun Kundu, Rupam Biswas, Linda Reinhard, Anirudha Dutta, Jochen Muellerdieckmann, M. S. Weiss, Somnath Mukherjee, Nishith Kumar Pal, Amit DasAbstract:Secretary proteins of Mycobacterium Tuberculosis are key players of the mycobacterial infection pathway. MTC28 is a 28-kDa proline-rich secretary antigen of Mycobacterium Tuberculosis and is only conserved in pathogenic strains of mycobacteria. Here we report the crystal structure of MTC28 at 2.8- and 2.15-A resolutions for the structure-based epitope design. MTC28 shares a "mog1p"-fold consisting of seven antiparallel β strands stacked between α helices. Five probable epitopes have been located on a solvent-accessible flexible region by computational analysis of the structure of MTC28. Simultaneously, the protein is digested with trypsin and the resulting fragments are purified by HPLC. Such 10 purified peptide fragments are screened against sera from patients infected with pulmonary Tuberculosis (PTB). Two of these 10 fragments, namely (127)ALDITLPMPPR(137) and (138)WTQVPDPNVPDAFVVIADR(156),are found to be major immunogenic epitopes that are localized on the outer surface of the protein molecule and are part of a single continuous epitope that have been predicted in silico Mutagenesis and antibody inhibition studies are in accordance with the results obtained from epitope mapping.
