An active site of growth hormone for eliciting the differentiation of preadipose 3T3-F442A cells to adipose cells.
In order to elucidate the active sites of growth hormone for eliciting the differentiation of preadipose 3T3-F442A cells to adipocytes, four artificial mutant variants of human growth hormone (hGH) modified in the loop region of amino acid residues 54-74 were prepared in Escherichia coli by site-directed mutagenesis. Although the P59A (replacement of Pro59 with Ala) variant retained almost the same biological- and receptor binding-activity as hGH, the P61A (replacement of Pro61 with Ala) and the P59A-P61A (replacement of both Pro59 and Pro61 with Ala) both exhibited about half the activity, and the delta (62-67) variant (deletion of the residues 62-67) exhibited only about 0.1% the activity of those of intact hGH. The results suggest that Pro61 may be involved in formation of the active conformation of hGH, but Pro59 may not, and that the amino acid residues around 62-67 may be critical for the specific biological features of hGH.
Uchida
E
E
National Institute of Hygienic Sciences, Tokyo, Japan.
Shimokawa
S
S
Nishida
Y
Y
Takasu
H
H
Ikehara
M
M
Uesugi
S
S
Tomita
K
K
Tanaka
A
A
Morikawa
M
M
Hayakawa
T
T
D000273:Adipose Tissue / Q000166:cytology* / Q000187:drug effects; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D013006:Growth Hormone / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000494:pharmacology; D013552:Swine
D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D013006:Growth Hormone