title: Osteogenic protein-1 (OP-1) expression and processing in Chinese hamster ovary cells: isolation of a soluble complex containing the mature and pro-domains of OP-1.
abstract: We have characterized the expression and processing of Osteogenic Protein-1 (hOP-1), a bone morphogenic protein of the TGF-beta family, in Chinese hamster ovary cells. The hOP-1 is initially synthesized as a monomeric 50 kDa pro-protein that is dimerized, glycosylated, and then proteolytically cleaved at the Arg-Xaa-Xaa-Arg maturation site in an acidic cellular compartment before secretion into the medium. Of the four potential N-linked glycosylation sites two are used, one in the mature domain and one in the pro-domain. Gel permeation chromatography of secreted hOP-1 in physiological buffers yields an apparent molecular weight of 110-120 k, indicating that after proteolytic processing the two pro-domains remain non-covalently associated with the disulfide linked mature dimer in a complex termed soluble hOP-1. Purified soluble hOP-1 is significantly more soluble in physiological buffers than the purified mature OP-1.
authors.affiliation: Creative BioMolecules, Hopkinton, Mass. 01748, USA.
authors.forename: W K
authors.initials: WK
authors.lastname: Jones
authors.forename: E A
authors.initials: EA
authors.lastname: Richmond
authors.forename: K
authors.initials: K
authors.lastname: White
authors.forename: H
authors.initials: H
authors.lastname: Sasak
authors.forename: W
authors.initials: W
authors.lastname: Kusmik
authors.forename: J
authors.initials: J
authors.lastname: Smart
authors.forename: H
authors.initials: H
authors.lastname: Oppermann
authors.forename: D C
authors.initials: DC
authors.lastname: Rueger
authors.forename: R F
authors.initials: RF
authors.lastname: Tucker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D016466:CHO Cells; D002850:Chromatography, Gel; D006224:Cricetinae; D006031:Glycosylation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014162:Transfection / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D011506:Proteins; D016212:Transforming Growth Factor beta