Covalent binding efficiency of the third and fourth complement proteins in relation to pH, nucleophilicity, and availability of hydroxyl groups.
The binding of [3H]glycerol and [3H]putrescine to C3 was studied in a fluid-phase system using trypsin as the C3 convertase. The binding of glycerol showed little variation in the pH range between 6.0 and 10.0. The binding of putrescine (pKa = 9.0) is rather ineffective below pH 7.5 but becomes more efficient as the pH of the reaction mixture increases. These results agree with the contention that the final step of the binding reaction is the transfer of the acyl group of the exposed thio ester of C3 to a nucleophile since the nucleophilicity of hydroxyl groups is rather independent of pH whereas only the unprotonated form of amino groups is nucleophilic. The inefficient reaction of amino groups with the exposed thio ester of C3 is also supported by the study of the inhibitory activity of serine and its two derivatives, N-acetylserine and O-methylserine, to the binding of [3H]glycerol to C3. N-Acetylserine showed an inhibitory activity equivalent to that of serine, whereas O-methylated serine showed only minimal activity. It can be concluded, therefore, that serine reacts with the thio ester of C3 by its hydroxyl group but not by its alpha-amino group. The ability of the alcohol group of various alkanes to inhibit the binding of [3H]glycerol to C3 was also studied. The primary alcohols inhibit the binding reaction with an efficiency that is similar to glycerol, and there are no significant differences in the binding efficiencies of methanol, ethanol, 1-propanol, and 1-butanol.(ABSTRACT TRUNCATED AT 250 WORDS)
D000438:Alcohols / Q000494:pharmacology; D003166:Complement Activating Enzymes / Q000378:metabolism; D003173:Complement C1s; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003179:Complement C3b / Q000378:metabolism*; D003181:Complement C4 / Q000378:metabolism*; D015935:Complement C4b; D005990:Glycerol / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008433:Mathematics; D011700:Putrescine / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D000438:Alcohols; D003181:Complement C4; D012694:Serine; D003179:Complement C3b; D015935:Complement C4b; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases; D014357:Trypsin; D003173:Complement C1s; D005990:Glycerol; D011700:Putrescine