Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core
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 glyco...
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Chemical Society (ACS)
2017
|
| Online Access: | http://hdl.handle.net/1721.1/106626 https://orcid.org/0000-0003-3761-3743 https://orcid.org/0000-0002-5749-7869 |
| _version_ | 1826206763482873856 |
|---|---|
| author | Luo, Lingqi Kozakov, Dima Vajda, Sandor Allen, Karen N. Lukose, Vinita Imperiali, Barbara |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Luo, Lingqi Kozakov, Dima Vajda, Sandor Allen, Karen N. Lukose, Vinita Imperiali, Barbara |
| author_sort | Luo, Lingqi |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T13:38:11Z |
| format | Article |
| id | mit-1721.1/106626 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:38:11Z |
| publishDate | 2017 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1066262022-09-28T15:12:45Z Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core Luo, Lingqi Kozakov, Dima Vajda, Sandor Allen, Karen N. Lukose, Vinita Imperiali, Barbara Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Imperiali, Barbara Lukose, Vinita Imperiali, Barbara 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. National Institutes of Health (U.S.) (NIH Grant Number: R21 AI101807) National Institutes of Health (U.S.) (NIH Grant Number: GM039334) National Institutes of Health (U.S.) (NIH Grant Number: R01 GM064700) National Institutes of Health (U.S.) (NIH Grant Number: R01 GM 061867) 2017-01-25T21:11:15Z 2017-01-25T21:11:15Z 2015-11 2015-11 Article http://purl.org/eprint/type/JournalArticle 0006-2960 1520-4995 http://hdl.handle.net/1721.1/106626 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. https://orcid.org/0000-0003-3761-3743 https://orcid.org/0000-0002-5749-7869 en_US http://dx.doi.org/10.1021/acs.biochem.5b01086 Biochemistry 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. application/pdf American Chemical Society (ACS) Prof. Imperiali via Courtney Crummett |
| spellingShingle | Luo, Lingqi Kozakov, Dima Vajda, Sandor Allen, Karen N. Lukose, Vinita Imperiali, Barbara Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title | Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title_full | Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title_fullStr | Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title_full_unstemmed | Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title_short | Conservation and Covariance in Small Bacterial Phosphoglycosyltransferases Identify the Functional Catalytic Core |
| title_sort | conservation and covariance in small bacterial phosphoglycosyltransferases identify the functional catalytic core |
| url | http://hdl.handle.net/1721.1/106626 https://orcid.org/0000-0003-3761-3743 https://orcid.org/0000-0002-5749-7869 |
| work_keys_str_mv | AT luolingqi conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore AT kozakovdima conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore AT vajdasandor conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore AT allenkarenn conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore AT lukosevinita conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore AT imperialibarbara conservationandcovarianceinsmallbacterialphosphoglycosyltransferasesidentifythefunctionalcatalyticcore |