title: Structural basis for the autoprocessing of zinc metalloproteases in the thermolysin family.
abstract: Thermolysin-like proteases (TLPs), a large group of zinc metalloproteases, are synthesized as inactive precursors. TLPs with a long propeptide (∼200 residues) undergo maturation following autoprocessing through an elusive molecular mechanism. We report the first two crystal structures for the autoprocessed complexes of a typical TLP, MCP-02. In the autoprocessed complex, Ala205 shifts upward by 33 Å from the previously covalently linked residue, His204, indicating that, following autocleavage of the peptide bond between His204 and Ala205, a large conformational change from the zymogen to the autoprocessed complex occurs. The eight N-terminal residues (residues Ala205-Gly212) of the catalytic domain form a new β-strand, nestling into two other β-strands. Simultaneously, the apparent T(m) of the autoprocessed complex increases 20 °C compared to that of the zymogen. The stepwise degradation of the propeptide begins with two sequential cuttings at Ser49-Val50 and Gly57-Leu58, which lead to the disassembly of the propeptide and the formation of mature MCP-02. Our findings give new insights into the molecular mechanism of TLP maturation.
authors.affiliation: State Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Shandong University, Jinan 250100, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.forename: Jue
authors.initials: J
authors.lastname: Wang
authors.forename: Da-Qi
authors.initials: DQ
authors.lastname: Yu
authors.forename: Fei
authors.initials: F
authors.lastname: Bian
authors.forename: Bin-Bin
authors.initials: BB
authors.lastname: Xie
authors.forename: Xiu-Lan
authors.initials: XL
authors.lastname: Chen
authors.forename: Bai-Cheng
authors.initials: BC
authors.lastname: Zhou
authors.forename: Lu-Hua
authors.initials: LH
authors.lastname: Lai
authors.forename: Zhi-Xin
authors.initials: ZX
authors.lastname: Wang
authors.forename: Jia-Wei
authors.initials: JW
authors.lastname: Wu
authors.forename: Yu-Zhong
authors.initials: YZ
authors.lastname: Zhang
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000235:genetics* / Q000502:physiology; D004792:Enzyme Precursors / Q000737:chemistry*; D008666:Metalloendopeptidases / Q000737:chemistry*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017422:Sequence Analysis, DNA
chemical_list: D004792:Enzyme Precursors; D008666:Metalloendopeptidases