title: A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry.
abstract: Currently protease assay kits, requiring substrate that is either radiolabeled or fluorescence labeled and specialized instruments, are all expensive. A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninhydrin-based method for assay of collagenase activity and its inhibitor screening using spectrophotometry. In the method, without multistep sample treatments and substrate labeling, the hydrolytic products were directly traced by ninhydrin. The method is expected to be suitable for not only the assay of collagenase activity but also the others matrix metalloproteinases activities, and can be used for kinetic study.
authors.affiliation: Department of Molecular Biology & Biochemistry, Xinxiang Medical University, Xinxiang, Henan 453003, People's Republic of China.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yun
authors.initials: Y
authors.lastname: Fu
authors.forename: Sufeng
authors.initials: S
authors.lastname: Zhou
authors.forename: Lixia
authors.initials: L
authors.lastname: Kang
authors.forename: Changzheng
authors.initials: C
authors.lastname: Li
mesh_terms: D017364:Collagenases / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical; D057075:Enzyme Assays / Q000379:methods*; D006868:Hydrolysis; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology*; D009555:Ninhydrin / Q000737:chemistry*; D010618:Phenanthrolines / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000737:chemistry; D013053:Spectrophotometry
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D010618:Phenanthrolines; D011092:Polyethylene Glycols; D017364:Collagenases; D009555:Ninhydrin