title: Determination of the structure form of the fourth ligand of zinc in Acutolysin A using combined quantum mechanical and molecular mechanical simulation.
abstract: Acutolysin A, which is isolated from the snake venom of Agkistrodon acutus, is a member of the SVMPs subfamily of the metzincin family, and it is a snake venom zinc metalloproteinase possessing only one catalytic domain. The catalytic zinc ion, in the active site, is coordinated in a tetrahedral manner with three imidazole nitrogen atoms of histidine and one oxygen atom. It is uncertain whether this oxygen atom is a water molecule or a hydroxide ion just from the three-dimensional X-ray crystal structure. The identity of the fourth ligand of zinc is theoretically determined for the first time by performing both combined quantum mechanical and molecular mechanical (QM/MM) simulation and high-level quantum mechanical calculations. All of the results obtained indicate that the fourth ligand in the active site of the reported X-ray crystal structure is a water molecule rather than a hydroxide anion. On the basis of these theoretical results, we note that the experimental observed pH dependence of the proteolytic and hemorrhagic activity of Acutolysin A can be attributed to the deprotonation of the zinc-bound water to yield a better nucleophile, the hydroxide ion. Structural analyses revealed structural details useful for the understanding of acutolysin catalytic mechanism.
authors.affiliation: Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, P. R. China.
authors.forename: Emilia L
authors.initials: EL
authors.lastname: Wu
authors.forename: Kin-Yiu
authors.initials: KY
authors.lastname: Wong
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.forename: Keli
authors.initials: K
authors.lastname: Han
authors.forename: Jiali
authors.initials: J
authors.lastname: Gao
mesh_terms: D020134:Catalytic Domain; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D006860:Hydrogen Bonding; D008666:Metalloendopeptidases / Q000737:chemistry*; D011487:Protein Conformation; D011789:Quantum Theory; D015032:Zinc / Q000737:chemistry*
chemical_list: D008666:Metalloendopeptidases; C115407:acutolysin A; D015032:Zinc