title: Cold-adapted maturation of thermophilic WF146 protease by mimicking the propeptide binding interactions of psychrophilic subtilisin S41.
abstract: Thermophilic WF146 protease matures efficiently at 60 degrees C, but quite slowly at low temperatures. In this report, seven amino acid residues involved in interactions between the mature domain and the propeptide of the enzyme were substituted by corresponding residues of psychrophilic subtilisin S41 to generate mutant Mut7 (S105G/G107D/Y117E/S136N/V143G/K144E/D145S). Mut3 (S105G/G107D/Y117E) and Mut4 (S136N/V143G/K144E/D145S) were also constructed. Transferring structural features from S41 endowed Mut7 with a remarkably increased maturation rate, as well as an improved caseinolytic activity at 25 degrees C. Moreover, Mut3 and Mut4 each obtained one of the above endowments. Further studies suggest that low-temperature activity and maturation rate are not necessarily linked, and uncoupling structural elements modulating the two properties may be advantageous to cold adaptation.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, PR China.
authors.forename: Yi-Ran
authors.initials: YR
authors.lastname: Yang
authors.forename: Hui
authors.initials: H
authors.lastname: Zhu
authors.forename: Nan
authors.initials: N
authors.lastname: Fang
authors.forename: Xiaoliang
authors.initials: X
authors.lastname: Liang
authors.forename: Chuan-Qi
authors.initials: CQ
authors.lastname: Zhong
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Tang
authors.forename: Ping
authors.initials: P
authors.lastname: Shen
authors.forename: Bing
authors.initials: B
authors.lastname: Tang
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D003080:Cold Temperature; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011489:Protein Denaturation; D013381:Subtilisins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D010455:Peptides; D013381:Subtilisins