Isolation and characterization of C1q, a subcomponent of the first component of complement, from human and rabbit sera.
1. C1q, a subcomponent of the first component of complement, has been isolated, in a haemolytically active and soluble form, by ion-exchange chromatography and gel filtration, from human and rabbit sera. Yields ranged from 10 to 25mg/litre of serum and the activity of final preparations was consistently in the range 5x10(3)-15x10(3) C1qH(50) units/mg. 2. The molecular weights of human and rabbit subcomponent C1q were 409600 and 417600, as determined by sedimentation equilibrium studies. 3. Subcomponent C1q from both species was shown to be composed of non-covalently linked subunits of approximately 57000 molecular weight as determined by gel-filtration or sedimentation equilibrium studies in 5.3m-guanidinium chloride. Reduction or oxidation of human and rabbit subcomponent C1q yielded three chains each having a molecular weight of approximately 23000 and which differed slightly in amino acid composition but markedly in carbohydrate content. The oxidized chains were separated, on a preparative scale, by ion-exchange chromatography in 8m-urea on DEAE-cellulose. 4. Both human and rabbit subcomponent C1q contained hydroxyproline, hydroxylysine, a high percentage of glycine and approximately 8% carbohydrate. Glutamic acid and aspartic acid were the free N-terminal amino acids of human subcomponent C1q whereas only serine was found in rabbit subcomponent C1q. 5. Collagenase digestion of human or rabbit subcomponent C1q caused a rapid loss of haemolytic activity which correlated with the breakdown of collagenous regions in the molecule.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001224:Aspartic Acid / Q000032:analysis; D001797:Blood Protein Electrophoresis; D002241:Carbohydrates / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003165:Complement System Proteins / Q000032:analysis* / Q000302:isolation & purification; D005971:Glutamates / Q000032:analysis; D005998:Glycine / Q000032:analysis; D006461:Hemolysis; D006801:Humans; D006901:Hydroxylysine / Q000032:analysis; D006909:Hydroxyproline / Q000032:analysis; D003012:Microbial Collagenase; D008970:Molecular Weight; D010084:Oxidation-Reduction; D011817:Rabbits; D012694:Serine / Q000032:analysis; D014461:Ultracentrifugation
D000596:Amino Acids; D002241:Carbohydrates; D005971:Glutamates; D006901:Hydroxylysine; D001224:Aspartic Acid; D012694:Serine; D003165:Complement System Proteins; D003012:Microbial Collagenase; D006909:Hydroxyproline; D005998:Glycine