Novel mechanism for carbamoyl-phosphate synthetase

A nucleotide switch for functionally equivalent domains

Michael Kothe, Binnur Eroglu, Holly Mazza, Henry Samudera, Susan Powers-Lee

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Carbamoyl-phosphate synthetases (CPSs) utilize two molecules of ATP at two internally duplicated domains, B and C. Domains B and C have recently been shown to be structurally [Thoden, J. B., Holden, H. M., Wesenberg, G., Raushel, F. M. and Rayment, I. (1997) Biochemistry 36, 6305-6316] and functionally [Guy, H. I. and Evans, D. R. (1996) J. Biol. Chem. 271,13762- 13769] equivalent. We have carried out a site-directed mutagenic analysis that is consistent with AlP binding to a palmate motif rather than to a Walker A/B motif in domains B and C. To accommodate our present findings, as well as the other recent findings of structural and functional equivalence, we are proposing a novel mechanism for CPS. In this mechanism utilization of ATP bound to domain C is coupled to carbamoyl-phosphate synthesis at domain B via a nucleotide switch, with the energy of ATP hydrolysis at domain C allowing domain B to cycle between two alternative conformations.

Original languageEnglish (US)
Pages (from-to)12348-12353
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number23
DOIs
StatePublished - Nov 11 1997
Externally publishedYes

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Carbamyl Phosphate
Ligases
Nucleotides
Adenosine Triphosphate
Biochemistry
Hydrolysis

ASJC Scopus subject areas

  • General

Cite this

Novel mechanism for carbamoyl-phosphate synthetase : A nucleotide switch for functionally equivalent domains. / Kothe, Michael; Eroglu, Binnur; Mazza, Holly; Samudera, Henry; Powers-Lee, Susan.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, No. 23, 11.11.1997, p. 12348-12353.

Research output: Contribution to journalArticle

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AB - Carbamoyl-phosphate synthetases (CPSs) utilize two molecules of ATP at two internally duplicated domains, B and C. Domains B and C have recently been shown to be structurally [Thoden, J. B., Holden, H. M., Wesenberg, G., Raushel, F. M. and Rayment, I. (1997) Biochemistry 36, 6305-6316] and functionally [Guy, H. I. and Evans, D. R. (1996) J. Biol. Chem. 271,13762- 13769] equivalent. We have carried out a site-directed mutagenic analysis that is consistent with AlP binding to a palmate motif rather than to a Walker A/B motif in domains B and C. To accommodate our present findings, as well as the other recent findings of structural and functional equivalence, we are proposing a novel mechanism for CPS. In this mechanism utilization of ATP bound to domain C is coupled to carbamoyl-phosphate synthesis at domain B via a nucleotide switch, with the energy of ATP hydrolysis at domain C allowing domain B to cycle between two alternative conformations.

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