Autolysis of a novel multidomain subtilase-cold-adapted deseasin MCP-01 is pH-dependent and the surface loops in its catalytic domain, the linker, and the P_proprotein domain are susceptible to proteolytic attack.
Cold-adapted deseasin MCP-01 is a novel type subtilase with a multidomain structure containing a catalytic domain, a linker, a P_proprotein domain, and a PKD domain. Its autolysis was pH-dependent due to its flexible structure. N-terminal sequence analysis of the autolytic peptides revealed four autolytic sites in the catalytic domain. Three of these are in the same loops as mesophilic subtilases and one is unlike anything previously reported. Two autolytic sites were deduced in its linker and three in its P_proprotein domain, indicating the linker and the P_proprotein domain are flexible and susceptible to proteolytic attacks. Therefore, during MCP-01 autolysis, the linker and the P_proprotein domain of MCP-01 were easily attacked by proteolysis, resulting in cleavage of the C-terminal region. At the same time, some autolytic sites in the surface loops of the catalytic domain were cleaved. This is the first report describing the autolytic mechanism of a multidomain subtilase.
Chen
Xiu-Lan
XL
The State Key Lab of Microbial Technology, Marine Biotechnology Research Center, Shandong University, Jinan 250100, China.
Zhang
Yu-Zhong
YZ
Lu
Jing-Tao
JT
Xie
Bin-Bin
BB
Sun
Cai-Yun
CY
Guo
Bing
B
D000595:Amino Acid Sequence; D001665:Binding Sites; D002021:Buffers; D020134:Catalytic Domain; D003080:Cold Temperature; D010450:Endopeptidases / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000737:chemistry*; D013499:Surface Properties; D013997:Time Factors
D002021:Buffers; D010455:Peptides; D010450:Endopeptidases; C477696:protease MCP-01, Pseudoaltermonas; D013381:Subtilisins