Localization of GAR transformylase in Escherichia coli and mammalian cells

Lata T. Gooljarsingh, Joseph Ramcharan, Simon Gilroy, Stephen Benkovic

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Enzymes of the de novo purine biosynthetic pathway may form a multienzyme complex to facilitate substrate flux through the ten serial steps constituting the pathway. One likely strategy for complex formation is the use of a structural scaffold such as the cytoskeletal network or subcellular membrane of the cell to mediate protein-protein interactions. To ascertain whether this strategy pertains to the de novo purine enzymes, the localization pattern of the third purine enzyme, glycinamide ribonucleotide transformylase (GAR Tfase) was monitored in live Escherichia coli and mammalian cells. Genes encoding human as well as E. coli GAR Tfase fused with green fluorescent protein (GFP) were introduced into their respective cells with regulated expression of proteins and localization patterns monitored by using confocal fluorescence microscopy. In both instances images showed proteins to be diffused throughout the cytoplasm. Thus, GAR Tfase is not localized to an existing cellular architecture, so this device is probably not used to concentrate the members of the pathway. However, discrete clusters of the pathway may still exist throughout the cytoplasm.

Original languageEnglish (US)
Pages (from-to)6565-6570
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number12
DOIs
StatePublished - Jun 5 2001

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Phosphoribosylglycinamide Formyltransferase
Escherichia coli
Cytoplasm
Proteins
Enzymes
Multienzyme Complexes
Biosynthetic Pathways
Green Fluorescent Proteins
Fluorescence Microscopy
Confocal Microscopy
Cell Membrane
Equipment and Supplies
Genes
purine

All Science Journal Classification (ASJC) codes

  • General

Cite this

Gooljarsingh, Lata T. ; Ramcharan, Joseph ; Gilroy, Simon ; Benkovic, Stephen. / Localization of GAR transformylase in Escherichia coli and mammalian cells. In: Proceedings of the National Academy of Sciences of the United States of America. 2001 ; Vol. 98, No. 12. pp. 6565-6570.
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Localization of GAR transformylase in Escherichia coli and mammalian cells. / Gooljarsingh, Lata T.; Ramcharan, Joseph; Gilroy, Simon; Benkovic, Stephen.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 12, 05.06.2001, p. 6565-6570.

Research output: Contribution to journalArticle

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