Intracellular proteolytic processing of the heavy chain of rat pre-alpha-inhibitor. The COOH-terminal propeptide is required for coupling to bikunin.
Pre-alpha-inhibitor is a serum protein consisting of two polypeptides named bikunin and heavy chain 3 (H3). Both polypeptides are synthesized in hepatocytes and while passing through the Golgi complex, bikunin, which carries a chondroitin sulfate chain, becomes covalently linked to the COOH-terminal amino acid residue of H3 via its polysaccharide. Immediately prior to this reaction, a COOH-terminal propeptide of 33 kDa is cleaved off from the heavy chain. Using COS-1 cells transfected with rat H3, we found that in the absence of bikunin, the cleaved propeptide remained bound to the heavy chain and that H3 lacking the propeptide sequence did not become linked to coexpressed bikunin. Sequencing of H3 secreted from COS-1 cells showed that part of the molecules had a 12-amino acid residue long NH2-terminal propeptide. Cleavage of this propeptide, which occurred in the endoplasmic reticulum, was found to require basic amino acid residues at P1, P2, and P6 suggesting that it is mediated by a Golgi enzyme in transit. Deletion of the NH2-terminal propeptide or blocking of its release affected neither transport nor coupling of the heavy chain to bikunin.
Department of Medical Biochemistry and Microbiology, University of Uppsala, Biomedical Center, Uppsala, Sweden. Thuveson@medkem.uu.se
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D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D006023:Glycoproteins / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017384:Sequence Deletion; D014360:Trypsin Inhibitor, Kunitz Soybean; D014361:Trypsin Inhibitors / Q000235:genetics / Q000378:metabolism*
D006023:Glycoproteins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011498:Protein Precursors; C109148:SPINT2 protein, human; D014361:Trypsin Inhibitors; C061151:pre-alpha-trypsin inhibitor; D014360:Trypsin Inhibitor, Kunitz Soybean