Correlation between the human and porcine complement system: a small-angle scattering study of cross immunity and methylamine-induced conformational changes of porcine C3 and C4 proteins.
The porcine complement proteins C3 and C4 have been isolated and then characterized using small-angle scattering methods. Within the limits of experimental errors, the porcine proteins are virtually identical with the corresponding human proteins as measured in terms of mol. wt, Mr and radius of gyration, R,: Mr(C3) = 198,000, Mr(C4) = 207,000, and R(C3) = 4.4 nm, R(C4) = 4.5 nm. The C3 and C4 proteins from pigs show cross-immunity with monoclonal antibodies (mAbs) raised against human C3 and C4, respectively. Using the Fab fragments of these mAbs as markers, it is indicated that porcine C3 and C4 undergo a conformational change after reaction with methylamine. The relatively large increase in the radius of gyration observed, delta R = 1.0-1.2 nm, going from the Fab complexes to the Fab complexes of the methylamine derivatives, is similar to that observed for human C3 under similar conditions. This may indicate that methylamine cleaves a labile thiol ester bond supposed to be present within the porcine proteins and that the epitopes interacting with the Fab fragments are very similar to those of the human proteins. Porcine C3 also resembles the human analogue by forming dimers after being subjected to methylamine and dilute lauryl sulphate: Mr = 404,000 and R = 7.9 nm.
Department of Chemistry, Swedish University of Agricultural Sciences, Uppsala.
R
R
Osterberg
B
B
Malmensten
T
T
Boive
U
U
Nilsson
T
T
Stigbrand
K
K
Mortensen
D000818:Animals; D000911:Antibodies, Monoclonal; D003176:Complement C3 / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D003181:Complement C4 / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D003429:Cross Reactions; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008744:Methylamines / Q000494:pharmacology*; D008968:Molecular Conformation; D012542:Scattering, Radiation; D013552:Swine / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003176:Complement C3; D003181:Complement C4; D007140:Immunoglobulin Fab Fragments; D008744:Methylamines; C027451:methylamine