A new colorimetric assay for methionyl aminopeptidases: Examination of the binding of a new class of pseudopeptide analog inhibitors Export

@article{citeulike:4002512, abstract = {A direct and convenient spectrophotometric assay has been developed for methionine aminopeptidases (MetAPs). The method employs the hydrolysis of a substrate that is a methionyl analogue of p -nitroaniline ( l -Met- p -NA), which releases the chromogenic product p -nitroaniline. This chromogenic product can be monitored continuously using a UV–Vis spectrophotometer set at 405 nm. The assay was tested with the type I MetAP from Escherichia coli ( Ec MetAP-I) and the type II MetAP from Pyrococcus furiosus ( Pf MetAP-II). Using l -Met- p -NA, the kinetic constants k cat and K m were determined for Ec MetAP-I and Pf MetAP-II and were compared with those obtained with a ” standard” high-performance liquid chromatography (HPLC) discontinuous assay. The assay has also been used to determine the temperature dependence of the kinetic constant k cat for Pf MetAP-II as well as to screen two novel pseudopeptide inhibitors of MetAPs. The results demonstrate that l -Met- p -NA provides a fast, convenient, and effective substrate for both type I and type II MetAPs and that this substrate can be used to quickly screen inhibitors of MetAPs.}, author = {Mitra, S. and Dygasholz, A. and Jiracek, J. and Zertova, M. and Zakova, L. and Holz, R.}, citeulike-article-id = {4002512}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.ab.2006.06.012}, citeulike-linkout-1 = {http://linkinghub.elsevier.com/retrieve/pii/S0003269706004192}, day = {01}, doi = {10.1016/j.ab.2006.06.012}, issn = {00032697}, journal = {Analytical Biochemistry}, keywords = {aminopeptidase, l-met-pna, methionyl}, month = {October}, number = {1}, pages = {43--49}, posted-at = {2009-02-03 21:49:57}, priority = {2}, title = {A new colorimetric assay for methionyl aminopeptidases: Examination of the binding of a new class of pseudopeptide analog inhibitors}, url = {http://dx.doi.org/10.1016/j.ab.2006.06.012}, volume = {357}, year = {2006} }

TY - JOUR ID - citeulike:4002512 L3 - citeulike-article-id:4002512 N2 - A direct and convenient spectrophotometric assay has been developed for methionine aminopeptidases (MetAPs). The method employs the hydrolysis of a substrate that is a methionyl analogue of p -nitroaniline ( l -Met- p -NA), which releases the chromogenic product p -nitroaniline. This chromogenic product can be monitored continuously using a UV–Vis spectrophotometer set at 405 nm. The assay was tested with the type I MetAP from Escherichia coli ( Ec MetAP-I) and the type II MetAP from Pyrococcus furiosus ( Pf MetAP-II). Using l -Met- p -NA, the kinetic constants k cat and K m were determined for Ec MetAP-I and Pf MetAP-II and were compared with those obtained with a “standard” high-performance liquid chromatography (HPLC) discontinuous assay. The assay has also been used to determine the temperature dependence of the kinetic constant k cat for Pf MetAP-II as well as to screen two novel pseudopeptide inhibitors of MetAPs. The results demonstrate that l -Met- p -NA provides a fast, convenient, and effective substrate for both type I and type II MetAPs and that this substrate can be used to quickly screen inhibitors of MetAPs. IS - 1 JF - Analytical Biochemistry SN - 00032697 EP - 49 TI - A new colorimetric assay for methionyl aminopeptidases: Examination of the binding of a new class of pseudopeptide analog inhibitors VL - 357 SP - 43 KW - aminopeptidase KW - l-met-pna KW - methionyl AU - Mitra, S AU - Dygasholz, A AU - Jiracek, J AU - Zertova, M AU - Zakova, L AU - Holz, R PY - 2006/10/01/ UR - http://dx.doi.org/10.1016/j.ab.2006.06.012 ER -