Characterization of the beta-chain N-terminus heterogeneity and the alpha-chain C-terminus of human platelet GPIIb. Posttranslational cleavage sites.
Human platelet glycoprotein IIb (GPIIb) and IIIa (GPIIIa) form a Ca2(+)-dependent heterodimer, the integrin GPIIb/IIIa, which functions as the fibrinogen receptor at the surface of activated platelets. GPIIB and GPIIIa are synthesized as single polypeptides from single messages and their amino acid sequences were derived from their cDNAs. The GPIIb precursor is proteolytically processed to yield the known disulphide-bonded two-chain (GPIIb alpha and GPIIb beta) covalent structure found in mature GPIIb. Our present protein chemical and mass spectrometric analyses indicate that the GPIIb precursor is proteolytically cleaved at two or three sites, to give rise to an homogeneous alpha-chain (GPIIb 1-856) single disulphide-bonded to one of the two beta-chains, which are present in a nearly 1:1 ratio: GPIIb beta 1 (860-1008), with pyroglutamic acid as its blocked N-terminal residue: and GPIIb beta 2 (872-1008), with the already known N-terminal sequence. These results satisfy the previously observed electrophoretic size-residue: and GPIIb beta 2 (872-1008), with the already known N-terminal sequence. These results satisfy the previously observed electrophoretic size-heterogeneity of the beta-chain, confirmed the potential cleavage sites in the junction region, and indicate a probable dual proteolytic processing of GPIIb, which may be relevant to the rest of the two-chain alpha-subunits of the integrin family.
Calvete
J J
JJ
Max Planck Institut für Biochemie, Martinsried, FRG.
Schäfer
W
W
Henschen
A
A
González-Rodríguez
J
J
D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D003488:Cyanogen Bromide; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D004220:Disulfides; D046911:Macromolecular Substances; D010980:Platelet Membrane Glycoproteins; D011498:Protein Precursors; D010450:Endopeptidases; D003488:Cyanogen Bromide; D002118:Calcium