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Publication Abstract from PubMed

Succinyl-CoA synthetase (SCS) from Thermus aquaticus was characterized biochemically via measurements of the activity of the enzyme and determination of its quaternary structure as well as its stability and refolding properties. The enzyme is most active between pH 8.0 and 8.4 and its activity increases with temperature to about 339 K. Gel-filtration chromatography and sedimentation equilibrium under native conditions demonstrated that the enzyme is a heterotetramer of two alpha-subunits and two beta-subunits. The activity assays showed that the enzyme uses either ADP/ATP or GDP/GTP, but prefers GDP/GTP. This contrasts with Escherichia coli SCS, which uses GDP/GTP but prefers ADP/ATP. To understand the nucleotide preference, T. aquaticus SCS was crystallized in the presence of GDP, leading to the determination of the structure in complex with GDP-Mn(2+). A water molecule and Pro20beta in T. aquaticus take the place of Gln20beta in pig GTP-specific SCS, interacting well with the guanine base and other residues of the nucleotide-binding site. This leads to the preference for GDP/GTP, but does not hinder the binding of ADP/ATP.

Biochemical and structural characterization of the GTP-preferring succinyl-CoA synthetase from Thermus aquaticus.,Joyce MA, Hayakawa K, Wolodko WT, Fraser ME Acta Crystallogr D Biol Crystallogr. 2012 Jul;68(Pt 7):751-62. doi:, 10.1107/S0907444912010852. Epub 2012 Jun 15. PMID:22751660^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.