Functional and structural characterization of a novel catechol-O-methyltransferase from Schizosaccharomyces pombe.
Catechol-O-methyltransferase (COMT1 ) catalyzes the transfer of a methyl group from S-adenosylmethionine (SAM) to various catechol substrates. COMTs play vital roles in physiological processes in animals, plants, and fungi, as well as bacteria, and have essential application values in industry. spCOMT is a probable COMT from Schizosaccharomyces pombe. It has an extraordinary intracellular distribution different from other homologs and would thus be predicted to perform a distinct physiological function. In this report, recombinant spCOMT was purified and kinetically characterized for the first time. The enzymology assays indicate that spCOMT is a metal-dependent enzyme and belongs to class I OMTs. In addition, the crystal structures of apo-spCOMT and SAM-complexed spCOMT were also presented, revealing that spCOMT possesses a conserved SAM-binding site and Mg2+ pocket, but a distinct substrate pocket was not present in homologs. The mutagenesis ITC analysis revealed the SAM recognition characteristics of spCOMT. Based on all of the above findings, we speculated about the putative substrates' characteristics and the substrate recognition mechanisms of spCOMT. This work will help in elucidating the physiological functions of spCOMT in S. pombe. © 2018 IUBMB Life, 71(3):330-339, 2019.
['Wang|Qing|Q|', 'Teng|Maikun|M|', 'Li|Xu|X|0000-0001-5221-0817']
D000595:Amino Acid Sequence D000818:Animals D020134:Catalytic Domain D002394:Catechol O-Methyltransferase D002396:Catechols D003001:Cloning, Molecular D018360:Crystallography, X-Ray D004926:Escherichia coli D005656:Fungal Proteins D015870:Gene Expression D005822:Genetic Vectors D006801:Humans D007700:Kinetics D051379:Mice D008958:Models, Molecular D009154:Mutation D011485:Protein Binding D000072756:Protein Conformation, alpha-Helical D000072757:Protein Conformation, beta-Strand D054730:Protein Interaction Domains and Motifs D011994:Recombinant Proteins D012436:S-Adenosylmethionine D012568:Schizosaccharomyces D016415:Sequence Alignment D040681:Structural Homology, Protein D013379:Substrate Specificity
catechol-O-methyltransferase Schizosaccharomyces pombe metal-dependent crystal structure SAM-dependent