Nature of the interaction between the C1q and C1r2S2 subunits of the first component of human complement.
The strength of interaction between the C1q and C1r2S2 subunits of C1 was studied as a function of temp. During centrifugation through sucrose density gradients at 4 degrees C, macromolecular C1 readily dissociated as it sedimented away from its free subunits. In contrast, at 20 degrees C, C1 remained associated as the 16S complex throughout centrifugation, thus indicating a stronger interaction between C1q and C1r2S2 at the higher temp. C1-inhibitor (C1-In) or nitrophenylguanidinobenzoate was present during centrifugation to prevent C1 activation. That native C1 was in fact the species being studied was confirmed by SDS-PAGE analysis. To investigate this temp dependence without using inhibitors, an alternative approach was used. Trace amounts of 125I-C1q were centrifuged through numerous sucrose density gradients, each of which contained a different concn of native C1r2S2 throughout the gradient. The s-rate of 125I-C1q increased with increasing C1r2S2 input. An association constant of 4.9 X 10(7) M-1 was calculated for this reversible interaction at 4 degrees C. However, at 20 degrees C, the data indicated a much higher affinity reaction since the addition of far less C1r2S2 was required for the s-rate of 125I-C1q to reach the 16S plateau. The presence of Cl-In did not affect these results. We have demonstrated that the association of C1q with C1r2S2 increases with increasing temp, a finding suggestive of a hydrophobic interaction. However, since we also show that C1 readily dissociates with increasing NaCl concn, the C1q-C1r2S2 interaction must, in fact, be ionic in nature. We therefore conclude that the temp dependence of the inter-subunit interaction is the result of a conformational change(s) within one of the subunits, and propose that this change may be similar to that occurring during Cl activation.
Ziccardi
R J
RJ
D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D003166:Complement Activating Enzymes; D015922:Complement C1q; D015923:Complement C1r; D003173:Complement C1s; D006801:Humans; D005779:Immunodiffusion; D009994:Osmolar Concentration; D013696:Temperature
D015922:Complement C1q; D003166:Complement Activating Enzymes; D015923:Complement C1r; D003173:Complement C1s