Ligand-Induced Structural Changes in Adenosine 5‘-Phosphosulfate Kinase from Penicillium chrysogenum,
- 25 October 2002
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 41 (46), 13672-13680
- https://doi.org/10.1021/bi026556b
Abstract
Adenosine 5‘-phosphosulfate (APS) kinase catalyzes the second reaction in the two-step, ATP-dependent conversion of inorganic sulfate to 3‘-phosphoadenosine 5‘-phosphosulfate (PAPS). PAPS serves as the sulfuryl donor for the biosynthesis of all sulfate esters and also as a precursor of reduced sulfur biomolecules in many organisms. Previously, we determined the crystal structure of ligand-free APS kinase from the filamentous fungus, Penicillium chrysogenum [MacRae et al. (2000) Biochemistry 39, 1613−1621]. That structure contained a protease-susceptible disordered region (“mobile lid”; residues 145−170). Addition of MgADP and APS, which together promote the formation of a nonproductive “dead-end” ternary complex, protected the lid from trypsin. This report presents the 1.43 Å resolution crystal structure of APS kinase with both ADP and APS bound at the active site and the 2.0 Å resolution structure of the enzyme with ADP alone bound. The mobile lid is ordered in both complexes and is shown to provide part of the binding site for APS. That site is formed primarily by the highly conserved Arg 66, Arg 80, and Phe 75 from the protein core and Phe 165 from the mobile lid. The two Phe residues straddle the adenine ring of bound APS. Arg 148, a completely conserved residue, is the only residue in the mobile lid that interacts directly with bound ADP. Ser 34, located in the apex of the P-loop, hydrogen-bonds to the 3‘-OH of APS, the phosphoryl transfer target. The structure of the binary E·ADP complex revealed further changes in the active site and N-terminal helix that occur upon the binding/release of (P)APS.Keywords
This publication has 13 references indexed in Scilit:
- Molecular cloning of a novel human PAPS synthetase which is differentially expressed in metastatic and non-metastatic colon carcinoma cellsThe International Journal of Biochemistry & Cell Biology, 1999
- Adenosine 5′-Phosphosulfate Kinase from Penicillium chrysogenumPublished by Elsevier BV ,1998
- Molecular Cloning, Expression, and Characterization of Human Bifunctional 3′-Phosphoadenosine 5′-Phosphosulfate Synthase and Its Functional DomainsPublished by Elsevier BV ,1998
- Biology and function of the reversible sulfation pathway catalysed by human sulfotransferases and sulfatasesChemico-Biological Interactions, 1998
- [20] Processing of X-ray diffraction data collected in oscillation modeMethods in Enzymology, 1997
- Crystal structures of rat acid phosphatase complexed with the transition‐state analogs vanadate and molybdateEuropean Journal of Biochemistry, 1994
- PROCHECK: a program to check the stereochemical quality of protein structuresJournal of Applied Crystallography, 1993
- Adenosine-5′-phosphosulfate kinase from Penicillium chrysogenum: Ligand binding properties and the mechanism of substrate inhibitionArchives of Biochemistry and Biophysics, 1991
- Adenosine-5′-phosphosulfate Kinase from Escherichia coli K12Journal of Biological Chemistry, 1989
- Stereochemical Criteria for Polypeptide and Protein Chain Conformations: II. Allowed Conformations for a Pair of Peptide UnitsBiophysical Journal, 1965