Protein synthesis in rabbit reticulocytes. A study of the mechanism of interreaction of fluorescently labeled co-eIF-2A with eIF-2 using fluorescence polarization.

P. Ghosh-Dastidar, D. Giblin, B. Yaghmai, A. Das, H. K. Das, Lawrence J Parkhurst, N. K. Gupta

Research output: Contribution to journalArticle

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Abstract

5-Dimethylaminonaphthalene-1-sulfonyl (dansyl)-Co-eIF-2A was prepared using homogeneous Co-eIF-2A. Dansyl-Co-eIF-2A was as active as untreated Co-eIF-2A when assayed for stimulation of ternary complex formation and also for protection of the ternary complex from dissociation by aurintricarboxylic acid. The mechanism of interaction of dansyl-Co-eIF-2A with eIF-2 was studied by measuring changes in fluorescence polarization. These studies indicate that dansyl-Co-eIF-2A interacts specifically with the ternary complex and does not interact with free eIF-2 or with two other high molecular weight protein complexes, Co-eIF-2B and Co-eIF-2C. Mg2+ inhibits ternary complex formation by eIF-2 and Co-eIF-2C relieves this Mg2+ inhibition of ternary complex formation. In both cases, the changes in fluorescence polarization of dansyl-Co-eIF-2A correlate well with the extent of ternary complex formed.

Original languageEnglish (US)
Pages (from-to)3826-3829
Number of pages4
JournalJournal of Biological Chemistry
Volume255
Issue number9
StatePublished - May 10 1980

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Fluorescence Polarization
Reticulocytes
Aurintricarboxylic Acid
Fluorescence
Polarization
Rabbits
Proteins
Molecular Weight
Molecular weight

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Protein synthesis in rabbit reticulocytes. A study of the mechanism of interreaction of fluorescently labeled co-eIF-2A with eIF-2 using fluorescence polarization. / Ghosh-Dastidar, P.; Giblin, D.; Yaghmai, B.; Das, A.; Das, H. K.; Parkhurst, Lawrence J; Gupta, N. K.

In: Journal of Biological Chemistry, Vol. 255, No. 9, 10.05.1980, p. 3826-3829.

Research output: Contribution to journalArticle

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AU - Ghosh-Dastidar, P.

AU - Giblin, D.

AU - Yaghmai, B.

AU - Das, A.

AU - Das, H. K.

AU - Parkhurst, Lawrence J

AU - Gupta, N. K.

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N2 - 5-Dimethylaminonaphthalene-1-sulfonyl (dansyl)-Co-eIF-2A was prepared using homogeneous Co-eIF-2A. Dansyl-Co-eIF-2A was as active as untreated Co-eIF-2A when assayed for stimulation of ternary complex formation and also for protection of the ternary complex from dissociation by aurintricarboxylic acid. The mechanism of interaction of dansyl-Co-eIF-2A with eIF-2 was studied by measuring changes in fluorescence polarization. These studies indicate that dansyl-Co-eIF-2A interacts specifically with the ternary complex and does not interact with free eIF-2 or with two other high molecular weight protein complexes, Co-eIF-2B and Co-eIF-2C. Mg2+ inhibits ternary complex formation by eIF-2 and Co-eIF-2C relieves this Mg2+ inhibition of ternary complex formation. In both cases, the changes in fluorescence polarization of dansyl-Co-eIF-2A correlate well with the extent of ternary complex formed.

AB - 5-Dimethylaminonaphthalene-1-sulfonyl (dansyl)-Co-eIF-2A was prepared using homogeneous Co-eIF-2A. Dansyl-Co-eIF-2A was as active as untreated Co-eIF-2A when assayed for stimulation of ternary complex formation and also for protection of the ternary complex from dissociation by aurintricarboxylic acid. The mechanism of interaction of dansyl-Co-eIF-2A with eIF-2 was studied by measuring changes in fluorescence polarization. These studies indicate that dansyl-Co-eIF-2A interacts specifically with the ternary complex and does not interact with free eIF-2 or with two other high molecular weight protein complexes, Co-eIF-2B and Co-eIF-2C. Mg2+ inhibits ternary complex formation by eIF-2 and Co-eIF-2C relieves this Mg2+ inhibition of ternary complex formation. In both cases, the changes in fluorescence polarization of dansyl-Co-eIF-2A correlate well with the extent of ternary complex formed.

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