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Alternate modes of binding in two crystal structures of alkaline phosphatase‐inhibitor complexes
Author(s) -
Holtz Kathleen M.,
Kantrowitz Evan R.,
Stec Boguslaw,
Myers Jason K.,
Antonelli Stephen M.,
Widlanski Theodore S.
Publication year - 2000
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.9.5.907
Subject(s) - phosphonate , chemistry , moiety , dissociation constant , crystallography , stereochemistry , hydrogen bond , crystal structure , alkaline phosphatase , binding site , enzyme , molecule , organic chemistry , biochemistry , receptor
Two high resolution crystal structures of Escherichia coli alkaline phosphatase (AP) in the presence of phosphonate inhibitors are reported. The phosphonate compounds, phosphonoacetic acid (PAA) and mercaptomethylphosphonic acid (MMP), bind competitively to AP with dissociation constants of 5.5 and 0.6 mM, respectively. The structures of the complexes of AP with PAA and MMP were refined at high resolution to crystallographic R ‐values of 19.0 and 17.5%, respectively. Refinement of the AP‐inhibitor complexes was carried out using X‐PLOR. The final round of refinement was done using SHELXL‐97. Crystallographic analyses of the inhibitor complexes reveal different binding modes for the two phosphonate compounds. The significant difference in binding constants can be attributed to these alternative binding modes observed in the high resolution X‐ray structures. The phosphinyl group of PAA coordinates to the active site zinc ions in a manner similar to the competitive inhibitor and product inorganic phosphate. In contrast, MMP binds with its phosphonate moiety directed toward solvent. Both enzyme‐inhibitor complexes exhibit close contacts, one of which has the chemical and geometrical potential to be considered an unconventional hydrogen bond of the type C‐H…X.