2VQR

Crystal structure of a phosphonate monoester hydrolase from rhizobium leguminosarum: a new member of the alkaline phosphatase superfamily

Summary for 2VQR

• Entry DOI: 10.2210/pdb2vqr/pdb
• Descriptor: PUTATIVE SULFATASE, MANGANESE (II) ION, CALCIUM ION, ... (6 entities in total)
• Functional Keywords: phosphonate monoester hydrolase, hydrolase, plasmid, formylglycine, phosphodiesterase, alkaline phosphatase superfamily
• Biological source: RHIZOBIUM LEGUMINOSARUM BV. VICIAE
• Total number of polymer chains: 1
• Total formula weight: 61334.52

Authors

Jonas, S.,Hyvonen, M.,Hollfelder, F. (deposition date: 2008-03-18, release date: 2008-09-30, Last modification date: 2024-11-13)

Primary citation

Jonas, S.,Van Loo, B.,Hyvonen, M.,Hollfelder, F.
A New Member of the Alkaline Phosphatase Superfamily with a Formylglycine Nucleophile: Structural and Kinetic Characterisation of a Phosphonate Monoester Hydrolase/Phosphodiesterase from Rhizobium Leguminosarum.
J.Mol.Biol., 384:120-, 2008

PubMed Abstract: The alkaline phosphatase superfamily comprises a large number of hydrolytic metalloenzymes such as phosphatases and sulfatases. We have characterised a new member of this superfamily, a phosphonate monoester hydrolase/phosphodiesterase from Rhizobium leguminosarum (R/PMH) both structurally and kinetically. The 1.42 A crystal structure shows structural homology to arylsulfatases with conservation of the core alpha/beta-fold, the mononuclear active site and most of the active-site residues. Sulfatases use a unique formylglycine nucleophile, formed by posttranslational modification of a cysteine/serine embedded in a signature sequence (C/S)XPXR. We provide mass spectrometric and mutational evidence that R/PMH is the first non-sulfatase enzyme shown to use a formylglycine as the catalytic nucleophile. R/PMH hydrolyses phosphonate monoesters and phosphate diesters with similar efficiency. Burst kinetics suggest that substrate hydrolysis proceeds via a double-displacement mechanism. Kinetic characterisation of active-site mutations establishes the catalytic contributions of individual residues. A mechanism for substrate hydrolysis is proposed on the basis of the kinetic data and structural comparisons with E. coli alkaline phosphatase and Pseudomonas aeruginosa arylsulfatase. R/PMH represents a further example of conservation of the overall structure and mechanism within the alkaline phosphatase superfamily.

PubMed: 18793651
DOI: 10.1016/J.JMB.2008.08.072

Experimental method

X-RAY DIFFRACTION (1.42 Å)