Identification of a sequence motif that confers SecB dependence on a SecB-independent secretory protein in vivo

Jinoh Kim, Debra A. Kendall

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

SecB is a cytosolic chaperone which facilitates the transport of a subset of proteins, including membrane proteins such as PhoE and Lamb and some periplasmic proteins such as maltose-binding protein, in Escherichia coli. However, not all proteins require SecB for transport, and proteins such as ribose-binding protein are exported efficiently even in SecB-null strains. The characteristics which confer SecB dependence on some proteins but not others have not been defined. To determine the sequence characteristics that are responsible for the SecB requirement, we have inserted a systematic series of short, polymeric sequences into the SecB independent protein alkaline phosphatase (PhoA). The extent to which these simple sequences convert alkaline phosphatase into a SecB-requiring protein was evaluated in vivo. Using this approach we have examined the roles of the polarity and charge of the sequence, as well as its location within the mature region, in conferring SecB dependence. We find that an insert with as few as 10 residues, of which 3 are basic, confers SecB dependence and that the mutant protein is efficiently exported in the presence of SecB. Remarkably, the basic motifs caused the protein to be translocated in a strict membrane potential-dependent fashion, indicating that the membrane potential is not a barrier to, but rather a requirement for, translocation of the motif. The alkaline phosphatase mutants most sensitive to the loss of SecB are those most sensitive to inhibition of SecA via azide treatment, consistent with the necessity for formation of a preprotein-SecB-SecA complex. Furthermore, the impact of the basic motif depends on location within the mature protein and parallels the accessibility of the location to the secretion apparatus.

Original languageEnglish (US)
Pages (from-to)1396-1401
Number of pages6
JournalJournal of Bacteriology
Volume180
Issue number6
StatePublished - Mar 1998
Externally publishedYes

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Alkaline Phosphatase
Proteins
Membrane Potentials
Carrier Proteins
Periplasmic Proteins
Maltose-Binding Proteins
Amino Acid Motifs
Ribose
Azides
Phosphoprotein Phosphatases
Mutant Proteins
Membrane Proteins
Escherichia coli

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Identification of a sequence motif that confers SecB dependence on a SecB-independent secretory protein in vivo. / Kim, Jinoh; Kendall, Debra A.

In: Journal of Bacteriology, Vol. 180, No. 6, 03.1998, p. 1396-1401.

Research output: Contribution to journalArticle

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