Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core
Author(s)
Luo, Lingqi; Kozakov, Dima; Vajda, Sandor; Allen, Karen N.; Lukose, Vinita; Imperiali, Barbara; ... Show more Show less![Thumbnail](/bitstream/handle/1721.1/106626/imperali%20LUKOSE-BIOCHEM-2016-FINAL.pdf.jpg?sequence=4&isAllowed=y)
Downloadimperali LUKOSE-BIOCHEM-2016-FINAL.pdf (18.73Mb)
PUBLISHER_POLICY
Publisher Policy
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Terms of use
Metadata
Show full item recordAbstract
Phosphoglycosyltransferases (PGTs) catalyze the transfer of a C1′-phosphosugar from a soluble sugar nucleotide diphosphate to a polyprenol phosphate. These enzymes act at the membrane interface, forming the first membrane-associated intermediates in the biosynthesis of cell-surface glycans and glycoconjugates, including glycoproteins, glycolipids, and the peptidoglycan in bacteria. PGTs vary greatly in both their membrane topologies and their substrate preferences. PGTs, such as MraY and WecA, are polytopic, while other families of uniquely prokaryotic enzymes have only a single predicted transmembrane helix. PglC, a PGT involved in the biosynthesis of N-linked glycoproteins in the enteropathogen Campylobacter jejuni, is representative of one of the structurally most simple members of the diverse family of small bacterial PGT enzymes. Herein, we apply bioinformatics and covariance-weighted distance constraints in geometry- and homology-based model building, together with mutational analysis, to investigate monotopic PGTs. The pool of 15000 sequences that are analyzed include the PglC-like enzymes, as well as sequences from two other related PGTs that contain a “PglC-like” domain embedded in their larger structures (namely, the bifunctional PglB family, typified by PglB from Neisseria gonorrheae, and WbaP-like enzymes, typified by WbaP from Salmonella enterica). Including these two subfamilies of PGTs in the analysis highlights key residues conserved across all three families of small bacterial PGTs. Mutagenesis analysis of these conserved residues provides further information about the essentiality of many of these residues in catalysis. Construction of a structural model of the cytosolic globular domain utilizing three-dimensional distance constraints, provided by conservation covariance analysis, provides additional insight into the catalytic core of these families of small bacterial PGT enzymes.
Date issued
2015-11Department
Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of ChemistryJournal
Biochemistry
Publisher
American Chemical Society (ACS)
Citation
Lukose, Vinita, Lingqi Luo, Dima Kozakov, Sandor Vajda, Karen N. Allen, and Barbara Imperiali. “Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core.” Biochemistry 54, no. 50 (December 22, 2015): 7326–7334.
Version: Author's final manuscript
ISSN
0006-2960
1520-4995