PurT-encoded glycinamide ribonucleotide transformylase. Accommodation of adenosine nucleotide analogs within the active site

James B. Thoden, Steven M. Firestine, Stephen Benkovic, Hazel M. Holden

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

PurT-encoded glycinamide ribonucleotide transformylase, or PurT transformylase, functions in purine biosynthesis by catalyzing the formylation of glycinamide ribonucleotide through a catalytic mechanism requiring Mg2+ATP and formate. From previous x-ray diffraction analyses, it has been demonstrated that PurT transformylase from Escherichia coli belongs to the ATP-grasp superfamily of enzymes, which are characterized by three structural motifs referred to as the A-, B-, and C-domains. In all of the ATP-grasp enzymes studied to date, the adenosine nucleotide ligands are invariably wedged between the B- and C-domains, and in some cases, such as biotin carboxylase and carbamoyl phosphate synthetase, the B-domains move significantly upon nucleotide binding. Here we present a systematic and high-resolution structural investigation of PurT transformylase complexed with various adenosine nucleotides or nucleotide analogs including Mg2+ATP, Mg2+-5′-adenylylimidodiphosphate, Mg2+-β,γ-methyleneadenosine 5′-triphosphate, Mg2+ATPγS, or Mg2+ADP. Taken together, these studies indicate that the conformation of the so-called "T-loop," delineated by Lys-155 to Gln-165, is highly sensitive to the chemical identity of the nucleotide situated in the binding pocket. This sensitivity to nucleotide identity is in sharp contrast to that observed for the "P-loop"-containing enzymes, in which the conformation of the binding motif is virtually unchanged in the presence or absence of nucleotides.

Original languageEnglish (US)
Pages (from-to)23898-23908
Number of pages11
JournalJournal of Biological Chemistry
Volume277
Issue number26
DOIs
StatePublished - Jun 28 2002

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Phosphoribosylglycinamide Formyltransferase
Adenosine
Catalytic Domain
Nucleotides
Hydroxymethyl and Formyl Transferases
Adenosine Triphosphate
formic acid
biotin carboxylase
Hand Strength
Conformations
Enzymes
Adenylyl Imidodiphosphate
Carbamyl Phosphate
Biosynthesis
Ligases
Adenosine Diphosphate
Escherichia coli
Diffraction
X-Rays
Ligands

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Thoden, James B. ; Firestine, Steven M. ; Benkovic, Stephen ; Holden, Hazel M. / PurT-encoded glycinamide ribonucleotide transformylase. Accommodation of adenosine nucleotide analogs within the active site. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 26. pp. 23898-23908.
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abstract = "PurT-encoded glycinamide ribonucleotide transformylase, or PurT transformylase, functions in purine biosynthesis by catalyzing the formylation of glycinamide ribonucleotide through a catalytic mechanism requiring Mg2+ATP and formate. From previous x-ray diffraction analyses, it has been demonstrated that PurT transformylase from Escherichia coli belongs to the ATP-grasp superfamily of enzymes, which are characterized by three structural motifs referred to as the A-, B-, and C-domains. In all of the ATP-grasp enzymes studied to date, the adenosine nucleotide ligands are invariably wedged between the B- and C-domains, and in some cases, such as biotin carboxylase and carbamoyl phosphate synthetase, the B-domains move significantly upon nucleotide binding. Here we present a systematic and high-resolution structural investigation of PurT transformylase complexed with various adenosine nucleotides or nucleotide analogs including Mg2+ATP, Mg2+-5′-adenylylimidodiphosphate, Mg2+-β,γ-methyleneadenosine 5′-triphosphate, Mg2+ATPγS, or Mg2+ADP. Taken together, these studies indicate that the conformation of the so-called {"}T-loop,{"} delineated by Lys-155 to Gln-165, is highly sensitive to the chemical identity of the nucleotide situated in the binding pocket. This sensitivity to nucleotide identity is in sharp contrast to that observed for the {"}P-loop{"}-containing enzymes, in which the conformation of the binding motif is virtually unchanged in the presence or absence of nucleotides.",
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PurT-encoded glycinamide ribonucleotide transformylase. Accommodation of adenosine nucleotide analogs within the active site. / Thoden, James B.; Firestine, Steven M.; Benkovic, Stephen; Holden, Hazel M.

In: Journal of Biological Chemistry, Vol. 277, No. 26, 28.06.2002, p. 23898-23908.

Research output: Contribution to journalArticle

TY - JOUR

T1 - PurT-encoded glycinamide ribonucleotide transformylase. Accommodation of adenosine nucleotide analogs within the active site

AU - Thoden, James B.

AU - Firestine, Steven M.

AU - Benkovic, Stephen

AU - Holden, Hazel M.

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N2 - PurT-encoded glycinamide ribonucleotide transformylase, or PurT transformylase, functions in purine biosynthesis by catalyzing the formylation of glycinamide ribonucleotide through a catalytic mechanism requiring Mg2+ATP and formate. From previous x-ray diffraction analyses, it has been demonstrated that PurT transformylase from Escherichia coli belongs to the ATP-grasp superfamily of enzymes, which are characterized by three structural motifs referred to as the A-, B-, and C-domains. In all of the ATP-grasp enzymes studied to date, the adenosine nucleotide ligands are invariably wedged between the B- and C-domains, and in some cases, such as biotin carboxylase and carbamoyl phosphate synthetase, the B-domains move significantly upon nucleotide binding. Here we present a systematic and high-resolution structural investigation of PurT transformylase complexed with various adenosine nucleotides or nucleotide analogs including Mg2+ATP, Mg2+-5′-adenylylimidodiphosphate, Mg2+-β,γ-methyleneadenosine 5′-triphosphate, Mg2+ATPγS, or Mg2+ADP. Taken together, these studies indicate that the conformation of the so-called "T-loop," delineated by Lys-155 to Gln-165, is highly sensitive to the chemical identity of the nucleotide situated in the binding pocket. This sensitivity to nucleotide identity is in sharp contrast to that observed for the "P-loop"-containing enzymes, in which the conformation of the binding motif is virtually unchanged in the presence or absence of nucleotides.

AB - PurT-encoded glycinamide ribonucleotide transformylase, or PurT transformylase, functions in purine biosynthesis by catalyzing the formylation of glycinamide ribonucleotide through a catalytic mechanism requiring Mg2+ATP and formate. From previous x-ray diffraction analyses, it has been demonstrated that PurT transformylase from Escherichia coli belongs to the ATP-grasp superfamily of enzymes, which are characterized by three structural motifs referred to as the A-, B-, and C-domains. In all of the ATP-grasp enzymes studied to date, the adenosine nucleotide ligands are invariably wedged between the B- and C-domains, and in some cases, such as biotin carboxylase and carbamoyl phosphate synthetase, the B-domains move significantly upon nucleotide binding. Here we present a systematic and high-resolution structural investigation of PurT transformylase complexed with various adenosine nucleotides or nucleotide analogs including Mg2+ATP, Mg2+-5′-adenylylimidodiphosphate, Mg2+-β,γ-methyleneadenosine 5′-triphosphate, Mg2+ATPγS, or Mg2+ADP. Taken together, these studies indicate that the conformation of the so-called "T-loop," delineated by Lys-155 to Gln-165, is highly sensitive to the chemical identity of the nucleotide situated in the binding pocket. This sensitivity to nucleotide identity is in sharp contrast to that observed for the "P-loop"-containing enzymes, in which the conformation of the binding motif is virtually unchanged in the presence or absence of nucleotides.

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