Application of robotics to steady state enzyme kinetics: analysis of tight-binding inhibitors of dipeptidyl peptidase IV

Using available commercial robotics and instrumentation, we developed a fully automated and rigorous steady state enzyme kinetic assay for dipeptidyl peptidase IV (DPP IV; E.C. 3.4.14.5). The automated assay was validated with isoleucyl thiazolidide, a potent inhibitor of DPP IV with K is=110 nM. Si...

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Veröffentlicht in:	Analytical biochemistry 2003-10, Vol.321 (2), p.157-166
Hauptverfasser:	Wang, Aiying, Huang, Yanting, Taunk, Prakash, Magnin, David R, Ghosh, Krishnendu, Robertson, James G
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Automation Binding, Competitive Competitive inhibitor Dipeptidyl Peptidase 4 - analysis Dipeptidyl Peptidase 4 - blood Dipeptidyl Peptidase 4 - metabolism Dipeptidyl peptidase IV Drug discovery Enzyme assay Humans Isoleucine - analogs & derivatives Isoleucine - metabolism Isoleucine - pharmacology Isoleucyl 4-cyanothiazolidide Kidney - enzymology Kinetics Models, Chemical Protease Inhibitors - metabolism Protease Inhibitors - pharmacology Rats Robotics - methods Signal window analysis Steady state kinetics Thiazoles - metabolism Thiazoles - pharmacology Tight-binding inhibitor
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