Adiponectin does not bind to gelatin: a new and easy way to purify high-molecular-weight adiponectin from human plasma.
Human plasma contains three forms of adiponectin, a trimer, a hexamer, and a high-molecular-weight (HMW) multimer. We previously reported HMW adiponectin was a gelatin-binding protein of 28 kDa (GBP28), it having been purified due to its affinity to gelatin-Cellulofine (Nakano, Y., et al. Isolation and characterization of GBP28, a novel gelatin-binding protein purified from human plasma. J. Biochem. 1996. 120: 803-12). Although HMW adiponectin binds to gelatin-Cellulofine, it cannot bind to gelatin-Sepharose. Gelatin-Cellulofine was made of formyl-Cellulofine and gelatin, and we found that HMW adiponectin binds to reduced formyl-Cellulofine with similar affinity as to gelatin-Cellulofine. Through only two steps using reduced formyl-Cellulofine and DEAE-Sepharose, HMW adiponectin can be effectively purified from human plasma.
Nakano
Yasuko
Y
Department of Pharmacogenomics School of Pharmacy, Showa University, 1-5-8, Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan. yanakano@pharm.showa-u.ac.jp
Shoji
Ayako
A
Arakawa
Atsushi
A
Iizuka
Yumiko
Y
Kikuchi
Yuriko
Y
Kobayashi
Maya
M
Tobe
Takashi
T
D052242:Adiponectin / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D005780:Gelatin / Q000737:chemistry*; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D010010:Osteoclasts / Q000187:drug effects; D053245:RANK Ligand / Q000494:pharmacology
C499918:ADIPOQ protein, human; D052242:Adiponectin; D053245:RANK Ligand; D005780:Gelatin; D002482:Cellulose