A recombinant-DNA-derived modification of human growth hormone (hGH44-191) with enhanced diabetogenic activity.
A modified human growth hormone (hGH) that lacks the first 43 residues of the intact hormone was prepared by recombinant-DNA technology. For preparative purposes an additional alanine was made the amino terminal residue. Sequence analysis and tryptic peptide mapping combined with amino acid analyses confirmed the structure of the polypeptide. Less than 2% N-terminal methionine was detected. The hGH44-191 was estimated to be at least 10 times more active than hGH in producing glucose intolerance in obese yellow mice (Avy/A) and was equipotent to hGH in increasing serum free fatty acids in fasted, hypophysectomized rats. The peptide did not promote growth in hypophysectomized rats nor did it exhibit early (1h) insulin-like activity in fasted, hypophysectomized rats, as indicated by its failure to lower blood glucose and fatty acids. The modified hGH was inactive in the Nb-2 cell assay but was about one-third as active as hGH in stimulating the pigeon crop sac. In radioimmunoassays using 125I-labeled hGH and polyclonal antibodies to intact hGH, cross-reactivity of hGH44-191 was less than 1%. We conclude that removal of the amino terminal portion of hGH enhances its diabetogenic properties, and that this activity does not depend upon the ability to promote growth. Furthermore, the insulin-like activity can be separated from its diabetogenic action by deletion of the first 43 amino terminal residues. This is the first report of a modified hGH that has anti-insulin effects greater than hGH itself.
Lutcher Brown Department of Biochemistry, Whittier Institute for Diabetes and Endocrinology, La Jolla, CA 92037.
U J
UJ
Lewis
L J
LJ
Lewis
M A
MA
Salem
N R
NR
Staten
S S
SS
Galosy
G G
GG
Krivi
D000595:Amino Acid Sequence; D000818:Animals; D010856:Columbidae; D003429:Cross Reactions; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced* / Q000378:metabolism; D005227:Fatty Acids / Q000097:blood*; D005813:Genes, Synthetic; D005947:Glucose / Q000378:metabolism; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000494:pharmacology / Q000633:toxicity*; D007016:Hypophysectomy; D008066:Lipolysis / Q000187:drug effects; D051379:Mice; D008820:Mice, Obese / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology / Q000633:toxicity*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology / Q000633:toxicity*; D015430:Weight Gain / Q000187:drug effects
D005227:Fatty Acids; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C071687:somatotropin (44-91); D013006:Growth Hormone; D005947:Glucose