Phylogeny of the third component of complement, C3: analysis of the conservation of human CR1, CR2, H, and B binding sites, concanavalin A binding sites, and thiolester bond in the C3 from different species.
The third component of complement, C3, binds to several other complement proteins. To study the diverse reactivities of C3, we analyzed the conservation of structural and functional features in the C3 from different species. First, we developed a method to purify swine (Po), rabbit (Rb), mouse (Mo), cobra (Co), Xenopus (Xe), axolotl (Ax), and trout (Tr) C3 from plasma. This involved protein precipitation by polyethylene glycol, followed by anion-exchange, gel filtration, and cation exchange chromatography. All C3's tested were comprised of two chains (alpha/beta-chain) and contain a thiolester bond within the alpha-chain. The two N-linked high-mannose carbohydrates found in human C3 were only conserved (as detected by ConA binding) in Rb C3. In contrast, Xe, Ax, and Tr C3 have this moiety only in the beta-chain and Po and Mo C3 only in the alpha-chain. Co C3, in contrast to cobra venom factor (CVF), lacks ConA binding carbohydrates in both chains. N-terminal amino acid sequence analysis of the alpha-, alpha'-, and beta-chains showed varying degrees of similarity within the different C3's. The N-termini of the Xe and Ax C3 beta-chains were found to be blocked. The conservation of binding sites in the different C3's for human complement receptors type one (CR1) and two (CR2) and for factors H and B was investigated due to the structural and functional similarities of these molecules and to the ability of some of them to bind to the same domain(s) in human C3.(ABSTRACT TRUNCATED AT 250 WORDS)
Alsenz
J
J
Basel Institute for Immunology, Switzerland.
Avila
D
D
Huemer
H P
HP
Esparza
I
I
Becherer
J D
JD
Kinoshita
T
T
Wang
Y
Y
Oppermann
S
S
Lambris
J D
JD
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003176:Complement C3 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D011415:Complement Factor B / Q000378:metabolism; D017242:Complement Factor H; D003208:Concanavalin A / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011414:Properdin / Q000378:metabolism; D011951:Receptors, Complement / Q000378:metabolism; D013045:Species Specificity; D014714:Vertebrates / Q000378:metabolism
C493495:CFH protein, human; D003176:Complement C3; D003180:Complement C3b Inactivator Proteins; D011951:Receptors, Complement; D011414:Properdin; D003208:Concanavalin A; D017242:Complement Factor H; D011415:Complement Factor B