Crystalline arrays of the Escherichia coli sn-glycerol-3-phosphate acyltransferase, an integral membrane protein

W. O. Wilkison, J. P. Walsh, J. M. Corless, R. M. Bell

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The gene encoding the Escherichia coli sn-glycerol-3-phosphate acyltransferase, plsB, was inserted into hybrid plasmids under transcriptional control of the λP(L) and tac promoters. Enzymatic activities 35-50-fold above wild type and a large increase in glycerol-P acyltransferase polypeptide were obtained. Thin section electron microscopy of the cells overproducing the glycerol-P acyltransferase revealed 235-245-Å diameter tubular structures associated with the cytoplasmic membrane. These structures were released from the cell by osmotic lysis and purified on Matrex Gel Green A. Subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that the dominant protein constituent of the tubules was the glycerol-P acyltransferase. Analysis of tubule-enriched fractions isolated by differential centrifugation revealed a decreased phospholipid to protein ratio as compared to total and cytoplasmic membrane fractions. At high magnification, negative stained tubules displayed ordered arrays of stain-excluding components projecting 50-60 Å from the cytoplasmic surface. Optical diffraction patterns from the micrographs contained intense layer lines at (1/78 Å) and (1/39 Å) along the tubule axis and a prominent spot at (1/62 Å) near the equator. From compositional and structural data, 18-37% of the polypeptide volume is estimated to lie within the hydrophobic domain of the tubule membrane.

Original languageEnglish (US)
Pages (from-to)9951-9958
Number of pages8
JournalJournal of Biological Chemistry
Volume261
Issue number21
StatePublished - 1986
Externally publishedYes

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Acyltransferases
Escherichia coli
Membrane Proteins
Glycerol
Crystalline materials
Membranes
Cell Membrane
Peptides
Gene encoding
Centrifugation
Electrophoresis
Sodium Dodecyl Sulfate
Diffraction patterns
Electron microscopy
Polyacrylamide Gel Electrophoresis
Phospholipids
Electron Microscopy
Proteins
Plasmids
Coloring Agents

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystalline arrays of the Escherichia coli sn-glycerol-3-phosphate acyltransferase, an integral membrane protein. / Wilkison, W. O.; Walsh, J. P.; Corless, J. M.; Bell, R. M.

In: Journal of Biological Chemistry, Vol. 261, No. 21, 1986, p. 9951-9958.

Research output: Contribution to journalArticle

Wilkison, W. O. ; Walsh, J. P. ; Corless, J. M. ; Bell, R. M. / Crystalline arrays of the Escherichia coli sn-glycerol-3-phosphate acyltransferase, an integral membrane protein. In: Journal of Biological Chemistry. 1986 ; Vol. 261, No. 21. pp. 9951-9958.
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