Modification of arginine residues in human growth hormone by 1,2-cyclohexanedione: effects on the binding capacity to lactogenic and somatogenic receptors.
Reactivity of arginine residues in human growth hormone was studied by reaction with 1,2-cyclohexanedione. Kinetic analysis of the data showed a good fit to a pseudo first order curve, with an apparent velocity constant k = 1.26 x 10(-2) min-1 and a maximum modification of 9.6 out of the 11 arginines of the molecule. Modification led to a decrease in binding capacity to both lactogenic and somatogenic rat liver receptors. In either case Tsou plots suggest that the modification of two arginine residues is responsible for this behavior, although it cannot be ascertained whether the two relevant residues are the same for both receptor types. Circular dichroism studies indicated no apparent changes in protein conformation in the modified hormone. Binding capacity was restored upon regeneration of arginines by incubation with Tris-HCl buffer. Only the carboxy-terminal peptide was isolated by HPLC from a tryptic digest of succinylated Arg-modified hGH, indicating that 183 is the nonreacting arginine residue.
Atlasovich
F M
FM
Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Caridad
J J
JJ
Nowicki
C
C
Santomé
J A
JA
Wolfenstein-Todel
C
C
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D003510:Cyclohexanes / Q000494:pharmacology*; D003512:Cyclohexanones / Q000378:metabolism / Q000494:pharmacology*; D013006:Growth Hormone / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism; D010928:Placental Lactogen / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018000:Receptors, Peptide; D011986:Receptors, Somatotropin / Q000378:metabolism*; D013329:Structure-Activity Relationship
D003510:Cyclohexanes; D003512:Cyclohexanones; D007202:Indicators and Reagents; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; D011986:Receptors, Somatotropin; C028202:placental lactogen receptor; C018031:1,2-cyclohexanedione; D013006:Growth Hormone; D010928:Placental Lactogen; D001120:Arginine