Application of "homogeneous assay for fluorescence concentrated on membrane" to the analysis of the substrate specificity of protease.
The utility of the homogeneous assay for fluorescence concentrated on membrane (HAFCOM) in the analysis of the substrate specificity of protease was investigated using tobacco etch virus (TEV) protease. The V(max) of TEV protease against variants of a substrate was obtained by a simple procedure. It was considered that HAFCOM was more accurate than other endpoint measurements of protease assay.
Tsuchida
Shirou
S
Department of Molecular Biosciences, School of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Japan. tsuchida@hoku-iryo-u.ac.jp
Shigematsu
Takashi
T
Yahara
Taemi
T
Nunome
Kana
K
Hamaue
Naoya
N
Aoki
Takashi
T
D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005453:Fluorescence; D013379:Substrate Specificity / Q000235:genetics
D010450:Endopeptidases; C099488:TEV protease