Cobamide structure depends on both lower ligand availability and CobT substrate specificity

Terence Spencer Crofts; Erica C. Seth; Amrita B. Hazra; Michiko E. Taga

doi:10.1016/j.chembiol.2013.08.006

Cobamide structure depends on both lower ligand availability and CobT substrate specificity

Terence Spencer Crofts, Erica C. Seth, Amrita B. Hazra, Michiko E. Taga^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

64 Scopus citations

Abstract

Cobamides are members of the vitamin B₁₂ family of cofactors that function in a variety of metabolic processes and are synthesized only by prokaryotes. Cobamides produced by different organisms vary in the structure of the lower axial ligand. Here we explore the molecular factors that control specificity in the incorporation of lower ligand bases into cobamides. We find that the cobT gene product, which activates lower ligand bases for attachment, limits the range of lower ligand bases that can be incorporated by bacteria. Furthermore, we demonstrate that the substrate specificity of CobT can be predictably altered by changing two active site residues. These results demonstrate that sequence variations in cobT homologs contribute to cobamide structural diversity. This analysis could open new routes to engineering specific cobamide production and understanding cobamide-dependent processes.

Original language	English (US)
Pages (from-to)	1265-1274
Number of pages	10
Journal	Chemistry and Biology
Volume	20
Issue number	10
DOIs	https://doi.org/10.1016/j.chembiol.2013.08.006
State	Published - Oct 24 2013

Funding

This work was supported by National Science Foundation grant MCB1122046 to M.E.T. We thank the members of the Taga laboratory and Sydney Kustu for constructive comments and members of the Taga laboratory for critical reading of this manuscript. We thank Kenny Mok for providing authentic phenolyl cobamide standards, Andrew Han for his help during phylogenetic analysis, and Shan Yi and Yujie Men for assistance with LC-MS/MS.

ASJC Scopus subject areas

Drug Discovery
Molecular Medicine
Molecular Biology
Biochemistry
Clinical Biochemistry
Pharmacology

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title = "Cobamide structure depends on both lower ligand availability and CobT substrate specificity",

abstract = "Cobamides are members of the vitamin B12 family of cofactors that function in a variety of metabolic processes and are synthesized only by prokaryotes. Cobamides produced by different organisms vary in the structure of the lower axial ligand. Here we explore the molecular factors that control specificity in the incorporation of lower ligand bases into cobamides. We find that the cobT gene product, which activates lower ligand bases for attachment, limits the range of lower ligand bases that can be incorporated by bacteria. Furthermore, we demonstrate that the substrate specificity of CobT can be predictably altered by changing two active site residues. These results demonstrate that sequence variations in cobT homologs contribute to cobamide structural diversity. This analysis could open new routes to engineering specific cobamide production and understanding cobamide-dependent processes.",

author = "Crofts, {Terence Spencer} and Seth, {Erica C.} and Hazra, {Amrita B.} and Taga, {Michiko E.}",

note = "Funding Information: This work was supported by National Science Foundation grant MCB1122046 to M.E.T. We thank the members of the Taga laboratory and Sydney Kustu for constructive comments and members of the Taga laboratory for critical reading of this manuscript. We thank Kenny Mok for providing authentic phenolyl cobamide standards, Andrew Han for his help during phylogenetic analysis, and Shan Yi and Yujie Men for assistance with LC-MS/MS. ",

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doi = "10.1016/j.chembiol.2013.08.006",

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T1 - Cobamide structure depends on both lower ligand availability and CobT substrate specificity

AU - Crofts, Terence Spencer

AU - Seth, Erica C.

AU - Hazra, Amrita B.

AU - Taga, Michiko E.

N1 - Funding Information: This work was supported by National Science Foundation grant MCB1122046 to M.E.T. We thank the members of the Taga laboratory and Sydney Kustu for constructive comments and members of the Taga laboratory for critical reading of this manuscript. We thank Kenny Mok for providing authentic phenolyl cobamide standards, Andrew Han for his help during phylogenetic analysis, and Shan Yi and Yujie Men for assistance with LC-MS/MS.

PY - 2013/10/24

Y1 - 2013/10/24

N2 - Cobamides are members of the vitamin B12 family of cofactors that function in a variety of metabolic processes and are synthesized only by prokaryotes. Cobamides produced by different organisms vary in the structure of the lower axial ligand. Here we explore the molecular factors that control specificity in the incorporation of lower ligand bases into cobamides. We find that the cobT gene product, which activates lower ligand bases for attachment, limits the range of lower ligand bases that can be incorporated by bacteria. Furthermore, we demonstrate that the substrate specificity of CobT can be predictably altered by changing two active site residues. These results demonstrate that sequence variations in cobT homologs contribute to cobamide structural diversity. This analysis could open new routes to engineering specific cobamide production and understanding cobamide-dependent processes.

AB - Cobamides are members of the vitamin B12 family of cofactors that function in a variety of metabolic processes and are synthesized only by prokaryotes. Cobamides produced by different organisms vary in the structure of the lower axial ligand. Here we explore the molecular factors that control specificity in the incorporation of lower ligand bases into cobamides. We find that the cobT gene product, which activates lower ligand bases for attachment, limits the range of lower ligand bases that can be incorporated by bacteria. Furthermore, we demonstrate that the substrate specificity of CobT can be predictably altered by changing two active site residues. These results demonstrate that sequence variations in cobT homologs contribute to cobamide structural diversity. This analysis could open new routes to engineering specific cobamide production and understanding cobamide-dependent processes.

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EP - 1274

JO - Chemistry and Biology

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ER -

Cobamide structure depends on both lower ligand availability and CobT substrate specificity

Abstract

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ASJC Scopus subject areas

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