Trapping and visualization of a covalent enzyme-phosphate intermediate.
Using a mutant version of E. coli alkaline phosphatase, we succeeded in trapping and determining the structure of the phospho-enzyme intermediate. The X-ray structure also revealed the catalytic water molecule, bound to one of the active site zinc ions, positioned ideally for the apical attack necessary for the hydrolysis of the intermediate.
Murphy
J E
JE
Stec
B
B
Ma
L
L
Kantrowitz
E R
ER
D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology; D006868:Hydrolysis; D008667:Metalloproteins / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D010710:Phosphates / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D014867:Water / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
D001426:Bacterial Proteins; D008667:Metalloproteins; D010710:Phosphates; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014867:Water; D000469:Alkaline Phosphatase; D015032:Zinc