Cloning and expression of adiponectin and its globular domain, and measurement of the biological activity in vivo.
3T3-L1-adipocytes produce the adipocyte complement related protein of 30 kD (ACRP30), which is exclusively expressed in differentiated adipocytes. Decreased expression of ACRP30 correlates with insulin resistance in mouse models of altered insulin sensitivity. Adiponectin, the human homologue of ACRP30, circulates in human plasma at high levels. Plasma adiponectin levels have been reported to be decreased in some insulin-resistant states, such as obesity and type II diabetes mellitus. Here, full-length adiponectin and its C-terminal globular head domain (gAdiponectin) were expressed in Escherichia coli and gAdiponectin was used to immunize a rabbit to obtain polyclonal antiserum with titer of 10,000. Adiponectin was detected in human plasma with the use of gAdiponectin anti-serum by Western blot analysis, which was also detected by gACRP30 anti-serum. Injection in alloxan-treated rats with purified recombinant fusion adiponectin or gAdiponectin transiently abolished hyperglycemia. So adiponectin and gAdiponectin might have activity as a glucose lowering agent and potentially as a therapeutic for metabolic disease. All these results suggested that the recombinant protein had biological activity, and provided a useful tool in further studies.
Hu
Xiao-Bo
XB
Research Center of Biotechnology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200233, China.
Zhang
Yu-Jian
YJ
Zhang
Hui-Tang
HT
Yang
Sheng-Li
SL
Gong
Yi
Y
D041721:3T3-L1 Cells; D052242:Adiponectin; D000496:Alloxan; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D001786:Blood Glucose / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000139:chemically induced / Q000188:drug therapy; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D011506:Proteins / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use
D052242:Adiponectin; D001786:Blood Glucose; D018076:DNA, Complementary; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D000496:Alloxan