Stereochemical and Kinetic Investigation of 32P-Labeled Inorganic Phosphate Exchange Reaction Catalyzed by Primer-Independent and Primer-Dependent Polynucleotide Phosphorylase from Micrococcus luteus

John F. Marlier, Floyd R. Bryant, Stephen Benkovic

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The SP diastereomer of adenosine 5'-O-(1-thiodiphosphate) (ADPαS) is a substrate for the 32P-labeled inorganic phosphate exchange reaction catalyzed by the T and I forms of polynucleotide phosphorylase. The exchange reaction occurs with retention of configuration. This exchange reaction is very slow when only ADPaS(5P) is present but is greatly activated by dinucleotide primers and ADPαS (RP), although the latter is not a substrate for the exchange reaction. Ap(S)A(RP) is an ~50% better activator of the exchange than the SP diastereomer. Furthermore, high levels of the ADPαS(SP) eliminate the activation by primers and by ADPαS (RP). A phosphatase activity is present with the I form of the enzyme which converts ADPaS(RP) to AMPS. This activity may be responsible for the formation of the 5'-phosphate end group for de novo polymerization or for the processivity of this reaction.

Original languageEnglish (US)
Pages (from-to)2212-2219
Number of pages8
JournalBiochemistry
Volume20
Issue number8
DOIs
StatePublished - Jan 1 1981

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Polyribonucleotide Nucleotidyltransferase
Micrococcus luteus
Phosphates
Depolymerization
Kinetics
Substrates
Phosphoric Monoester Hydrolases
Polymerization
Chemical activation
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

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title = "Stereochemical and Kinetic Investigation of 32P-Labeled Inorganic Phosphate Exchange Reaction Catalyzed by Primer-Independent and Primer-Dependent Polynucleotide Phosphorylase from Micrococcus luteus",
abstract = "The SP diastereomer of adenosine 5'-O-(1-thiodiphosphate) (ADPαS) is a substrate for the 32P-labeled inorganic phosphate exchange reaction catalyzed by the T and I forms of polynucleotide phosphorylase. The exchange reaction occurs with retention of configuration. This exchange reaction is very slow when only ADPaS(5P) is present but is greatly activated by dinucleotide primers and ADPαS (RP), although the latter is not a substrate for the exchange reaction. Ap(S)A(RP) is an ~50{\%} better activator of the exchange than the SP diastereomer. Furthermore, high levels of the ADPαS(SP) eliminate the activation by primers and by ADPαS (RP). A phosphatase activity is present with the I form of the enzyme which converts ADPaS(RP) to AMPS. This activity may be responsible for the formation of the 5'-phosphate end group for de novo polymerization or for the processivity of this reaction.",
author = "Marlier, {John F.} and Bryant, {Floyd R.} and Stephen Benkovic",
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Stereochemical and Kinetic Investigation of 32P-Labeled Inorganic Phosphate Exchange Reaction Catalyzed by Primer-Independent and Primer-Dependent Polynucleotide Phosphorylase from Micrococcus luteus. / Marlier, John F.; Bryant, Floyd R.; Benkovic, Stephen.

In: Biochemistry, Vol. 20, No. 8, 01.01.1981, p. 2212-2219.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Stereochemical and Kinetic Investigation of 32P-Labeled Inorganic Phosphate Exchange Reaction Catalyzed by Primer-Independent and Primer-Dependent Polynucleotide Phosphorylase from Micrococcus luteus

AU - Marlier, John F.

AU - Bryant, Floyd R.

AU - Benkovic, Stephen

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N2 - The SP diastereomer of adenosine 5'-O-(1-thiodiphosphate) (ADPαS) is a substrate for the 32P-labeled inorganic phosphate exchange reaction catalyzed by the T and I forms of polynucleotide phosphorylase. The exchange reaction occurs with retention of configuration. This exchange reaction is very slow when only ADPaS(5P) is present but is greatly activated by dinucleotide primers and ADPαS (RP), although the latter is not a substrate for the exchange reaction. Ap(S)A(RP) is an ~50% better activator of the exchange than the SP diastereomer. Furthermore, high levels of the ADPαS(SP) eliminate the activation by primers and by ADPαS (RP). A phosphatase activity is present with the I form of the enzyme which converts ADPaS(RP) to AMPS. This activity may be responsible for the formation of the 5'-phosphate end group for de novo polymerization or for the processivity of this reaction.

AB - The SP diastereomer of adenosine 5'-O-(1-thiodiphosphate) (ADPαS) is a substrate for the 32P-labeled inorganic phosphate exchange reaction catalyzed by the T and I forms of polynucleotide phosphorylase. The exchange reaction occurs with retention of configuration. This exchange reaction is very slow when only ADPaS(5P) is present but is greatly activated by dinucleotide primers and ADPαS (RP), although the latter is not a substrate for the exchange reaction. Ap(S)A(RP) is an ~50% better activator of the exchange than the SP diastereomer. Furthermore, high levels of the ADPαS(SP) eliminate the activation by primers and by ADPαS (RP). A phosphatase activity is present with the I form of the enzyme which converts ADPaS(RP) to AMPS. This activity may be responsible for the formation of the 5'-phosphate end group for de novo polymerization or for the processivity of this reaction.

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