Characterization of the C-terminal propeptide involved in bacterial wall spanning of alpha-amylase from the psychrophile Alteromonas haloplanctis

J Biol Chem. 1998 May 15;273(20):12109-15. doi: 10.1074/jbc.273.20.12109.

Abstract

The antarctic psychrophile Alteromonas haloplanctis secretes a Ca2+- and Cl--dependent alpha-amylase. The nucleotide sequence of the amy gene and the amino acid sequences of the gene products indicate that the alpha-amylase precursor is a preproenzyme composed by the signal peptide (24 residues), the mature alpha-amylase (453 residues, 49 kDa), and a long C-terminal propeptide or secretion helper (192 residues, 21 kDa). In cultures of the wild-type strain, the 70-kDa precursor is secreted at the mid-exponential phase and is cleaved by a nonspecific protease into the mature enzyme and the propeptide. The purified C-terminal propeptide displays several features common to beta-pleated transmembrane proteins. It has no intramolecular chaperone function because active alpha-amylase is expressed by Escherichia coli in the absence of the propeptide coding region. In E. coli, the 70-kDa precursor is directed toward the supernatant. When the alpha-amylase coding region is excised from the gene, the secretion helper can still promote its own membrane spanning. It can also accept a foreign passenger, as shown by the extracellular routing of a beta-lactamase-propeptide fusion protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins*
  • Base Sequence
  • Cell Wall / enzymology*
  • Circular Dichroism
  • DNA, Bacterial
  • Enzyme Precursors / metabolism
  • Escherichia coli / genetics
  • Gram-Negative Bacteria / enzymology*
  • Hydrolysis
  • Molecular Sequence Data
  • Mutagenesis
  • Protein Folding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spectrophotometry, Ultraviolet
  • alpha-Amylases / chemistry
  • alpha-Amylases / genetics
  • alpha-Amylases / metabolism*
  • beta-Lactamases / metabolism

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Enzyme Precursors
  • Recombinant Proteins
  • alpha-Amylases
  • alpha-amylase carboxy terminal propeptide, Pseudoalteromonas haloplanktis
  • beta-Lactamases

Associated data

  • GENBANK/X58627