Binding of radioiodinated human beta-endorphin to serum proteins from rats and humans, determined by several methods.
Binding of immunoreactive radioiodinated human beta-endorphin (125I-beta-EP) to rat serum was demonstrated by gel filtration of 125I-beta-EP in pooled rat serum on Sephadex G-200. Two radioactive peaks associated with proteins eluted from the column. The first peak eluted at the void volume containing lipoproteins, alpha 2- and beta 2-macroglobulins, and the second peak at the fraction of albumin. Binding of 125I-beta-EP to albumin was directly proved by gel filtration of 125I-beta-EP in buffer containing 4% human serum albumin on Sephadex G-200. Equilibrium dialysis was not applicable to investigating the interaction of 125I-beta-EP with serum proteins, because of the intense nonspecific adsorption to the semipermeable membrane and the degradation of the peptide during dialysis. Therefore, in order to quantitatively evaluate the binding of 125I-beta-EP in sera from rats and humans, we utilized four other methods (ultrafiltration, charcoal adsorption, polyethylene glycol precipitation and equilibrium gel filtration). These methods corresponded well with each other and indicated 35-44% binding of 125I-beta-EP in rat serum. Binding of 125I-beta-EP in normal human serum was 36%, determined by ultrafiltration. Serum protein binding of 125I-beta-EP was concentration independent over the concentration range studied (1-1000 nM).
Sato
H
H
Sugiyama
Y
Y
Sawada
Y
Y
Iga
T
T
Hanano
M
M
D000327:Adsorption; D000818:Animals; D001798:Blood Proteins / Q000378:metabolism*; D002606:Charcoal; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D004723:Endorphins / Q000378:metabolism*; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism; D008259:Macroglobulins / Q000378:metabolism; D008297:Male; D011092:Polyethylene Glycols; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D012709:Serum Albumin / Q000378:metabolism; D014462:Ultrafiltration; D001615:beta-Endorphin
D001798:Blood Proteins; D004723:Endorphins; D008074:Lipoproteins; D008259:Macroglobulins; D012709:Serum Albumin; D002606:Charcoal; D011092:Polyethylene Glycols; D001615:beta-Endorphin