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Media type:
E-Article
Title:
Nascent membrane and presecretory proteins synthesized in Escherichia coli associate with signal recognition particle and trigger factor
Contributor:
Valent, Quido A.;
De Gier, Jan‐Willem L.;
Heijne, Gunnar von;
Kendall, Debra A.;
Ten Hagen‐Jongman, Corinne M.;
Oudega, Bauke;
Luirink, Joen
imprint:
Wiley, 1997
Published in:Molecular Microbiology
Language:
English
DOI:
10.1046/j.1365-2958.1997.4431808.x
ISSN:
0950-382X;
1365-2958
Origination:
Footnote:
Description:
<jats:p>The <jats:italic>Escherichia coli</jats:italic> signal recognition particle (SRP) and trigger factor are cytoplasmic factors that interact with short nascent polypeptides of presecretory and membrane proteins produced in a heterologous <jats:italic>in vitro</jats:italic> translation system. In this study, we use an <jats:italic>E. coli in vitro</jats:italic> translation system in combination with bifunctional cross‐linking reagents to investigate these interactions in more detail in a homologous environment. Using this approach, the direct interaction of SRP with nascent polypeptides that expose particularly hydrophobic targeting signals is demonstrated, suggesting that inner membrane proteins are the primary physiological substrate of the <jats:italic>E. coli</jats:italic> SRP. Evidence is presented that the overproduction of proteins that expose hydrophobic polypeptide stretches, titrates SRP. In addition, trigger factor is efficiently cross‐linked to nascent polypeptides of different length and nature, some as short as 57 amino acid residues, indicating that it is positioned near the nascent chain exit site on the <jats:italic>E. coli</jats:italic> ribosome.</jats:p>