The fourth component of human complement treated with amines or chaotropes or frozen-thawed (C4b-like C4): interaction with C4 binding protein and cleavage by C3b/C4b inactivator.
We have shown previously that C4 treated with amines or chaotropes, although uncleaved, exhibits properties that are similar to C4b. Studies by other groups suggest that this C4b-like form of C4 is characterized by the lack of an internal thiolester bond that is present in native C4. We report here that C4 treated with N2H4 or KSCN or frozen-thawed, unlike native C4, forms a complex with C4-binding protein (C4-bp) and is cleaved by C3b/C4b inactivator (I). Fragmentation of C4b-like C4 by I occurs without previous cleavage to C4b and requires the presence of C4-bp. Cleavage of C4b-like C4 proceeds in two steps: a small fragment (16,000 m.w.) is released first, followed by cleavage of the remaining alpha-chain fragment (83,000 m.w.) into polypeptides of 46,000 (C4d) and 32,000 m.w. All fragments, except the 46,000 m.w. fragment, are disulfide-linked to the beta- and/or gamma-chains of C4. Cleavage of C4b-like C4 probably occurs at the same points in the alp a-chain as in C4b; however, C4b-like c4 also contains C4a. Based on the m.w. determinations, the C4a portion of the alpha-chain is present in the 83,000 m.w. fragment, and after the second cleavage, in the 32,000 m.w. fragment of C4b-like C4. These findings suggest the following alignment of alpha-chain fragments: the N-terminal C4a portion is attached to a polypeptide of about 25,000 m.w. (alpha 3), which is followed in the sequence by C4d (alpha 2) (46,000 m.w.) and a small 16,000 m.w. fragment (alpha 4) that forms the C-terminus.
D000588:Amines / Q000494:pharmacology*; D001798:Blood Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D003181:Complement C4 / Q000378:metabolism*; D015935:Complement C4b; D003169:Complement Inactivator Proteins; D005615:Freezing; D006023:Glycoproteins; D006801:Humans; D007460:Iodoacetamide / Q000378:metabolism; D013861:Thiocyanates / Q000494:pharmacology*; D013997:Time Factors
D000588:Amines; D001798:Blood Proteins; D002352:Carrier Proteins; D003180:Complement C3b Inactivator Proteins; D003181:Complement C4; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D013861:Thiocyanates; D015935:Complement C4b; C009941:potassium thiocyanate; D007460:Iodoacetamide