Selective purification and characterization of adiponectin multimer species from human plasma.
Adiponectin is an adipocyte-derived hormone and known to form several species of multimer, however, the precise components of each multimer have not been fully determined. We purified each multimer adiponectin selectively from human plasma and characterized them by affinity columns using anti-adiponectin, gelatin, or anti-albumin antibody and gel filtration. We found that adiponectin exists as four species of multimers in human plasma. According to their migrating mobility and N-terminal amino acid analysis, we defined them as a trimer, albumin-binding trimer, hexamer, and HMW. Low pH shifted HMW to hexamer, raising the possibility that HMW is a 12 mer or larger multimer. We also showed that HMW had the highest binding activity to the membrane fractions of C2C12 myocytes and activated AMPK most potently. Our results indicate that adiponectin forms diverse multimer species and at least some of the functional properties are dependent on a multimer status.
Hada
Yusuke
Y
Department of Metabolic Diseases, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan.
Yamauchi
Toshimasa
T
Waki
Hironori
H
Tsuchida
Atsushi
A
Hara
Kazuo
K
Yago
Hirokazu
H
Miyazaki
Osamu
O
Ebinuma
Hiroyuki
H
Kadowaki
Takashi
T
D055372:AMP-Activated Protein Kinases; D017667:Adipocytes / Q000737:chemistry*; D052242:Adiponectin / Q000737:chemistry / Q000302:isolation & purification*; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism; D046912:Multiprotein Complexes; D010949:Plasma / Q000737:chemistry*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism
D052242:Adiponectin; D009097:Multienzyme Complexes; D046912:Multiprotein Complexes; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases