The subtilisin Carlsberg pro-region is a membrane anchorage for two fusion proteins produced in Bacillus subtilis.
The extracellular serylprotease subtilisin Carlsberg (SubC) of Bacillus licheniformis is produced in a precursor form which includes a signal peptide (sp) and a pro-region. We have constructed a fusion protein in which the sp, pro-region and 38 amino acids (aa) at the N terminus of SubC were joined to the immunoglobulin (Ig) G-binding protein G produced by group G streptococci. The fused SubC::protein G was purified on IgG-Sepharose. IgG-binding material derived from membrane or supernatant fractions had different N termini, indicating that release from the membrane occurred only after removal of the pro-region. The proteolytic pattern was identical when SubC::protein G was produced in Bacillus subtilis 168 wild type or in a protease-deficient strain. The sp cleavage point was also defined in the membrane-derived material.
Egnell
P
P
Center for BioTechnology, Karolinska Institute, Huddinge, Sweden.
Flock
J I
JI
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D007074:Immunoglobulin G / Q000378:metabolism; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D013381:Subtilisins