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Related Experiment Videos

Bacteriophage M13 procoat protein inserts into the plasma membrane as a loop structure.

A Kuhn1

  • 1Microbiology Department, University of Basel, Switzerland.

Science (New York, N.Y.)
|December 4, 1987
PubMed
Summary

Bacteriophage M13 procoat inserts into the bacterial membrane as a loop. The leader peptide

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Area of Science:

  • Molecular Biology
  • Bacteriophage Biology
  • Membrane Protein Insertion

Background:

  • The major coat protein of bacteriophage M13 is synthesized as a precursor, procoat.
  • Procoat contains a 23-residue N-terminal leader sequence essential for membrane insertion.

Purpose of the Study:

  • To investigate the membrane insertion mechanism of bacteriophage M13 procoat.
  • To determine the topology of the leader peptide during membrane insertion.

Main Methods:

  • Construction of a fusion protein: M13 procoat fused to cytoplasmic ribulokinase.
  • Expression of the fusion protein in Escherichia coli.
  • Analysis of protein processing and topology using leader peptidase and protease protection assays.

Main Results:

  • The fusion protein inserts into the Escherichia coli plasma membrane.
  • Leader peptidase processes the fusion protein into a leader peptide and mature coat protein.
  • The N-terminus of the leader peptide remains in the cytoplasm, protected from external protease.

Conclusions:

  • Bacteriophage M13 procoat inserts into the membrane via a loop structure.
  • The N-terminus of the M13 procoat leader peptide remains in the cytoplasm during membrane insertion.

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