Carboxylate groups in bovine somatotropin involved in growth promoting activity. Theoretical models based upon individual kinetic constants to interpret the activity decay after chemical modification

Modification of approximately one fifth of the carboxylate groups in bovine somatotropin with a water soluble carbodiimide caused loss of growth promoting potency pointing to the existence of residues related to the hormonal activity among those belonging to a fast reacting set. A sigmoidal curve was obtained whether the inactivation process was referred to reaction time or degree of modification. Isoelectrofocusing of derivatives released the native hormone from responsibility for the biological potency exerted by preparations with 1.5–2.6 modified carboxylate groups. Examination of the individual reaction kinetics of the 11 fast reacting residues, in turn, excluded the possibility of the sigmoidal character of the inactivation curve being caused by a nonexponential disappearance of essential residues, as a possible consequence of the chemical modification of others. According to synthetic models, the experimental curve may be the consequence of the effect of cumulative modification of 2 or 3 out of a set of 3 to 8 relevant residues.