Molecular forms of cholecystokinin in plasma from normal and gastrectomized human subjects following a fat meal

Abstract

The present study was undertaken to characterize molecular forms of cholecystokinin (CCK) in human fat-stimulated plasma by Sephadex G50 column chromatography followed by radioimmunoassays employing 3 different region-specific antibodies. CCK was extracted and concentrated from plasma of healthy subjects by adsorption to SEP-PAK C18 cartridges and from plasma of gastrectomized patients by addition of 96% ethanol. Antibody 1703 binds to carboxy-terminal CCK-peptides containing at least 14 amino acid residues, antibody T204 to sulfated carboxy-terminal CCK-peptides and antibody 5135 to carboxy-terminal forms of CCK and gastrin. Four molecular forms of CCK were consistently demonstrated; peak I eluted in the void volume and comprised 1.8-10.2% of CCK-immunoreactivity, peak II eluted between the void volume and the CCK-33/39 standard and comprised 9.8-21.6%, peak III eluted at the position of the CCK-33/39 standard and comprised 42.4-55.4%, and peak IV eluted between the CCK-33/39 and CCK-14 standards and comprised 25.4-40.1% of CCK immunoreactivity. Since these 4 molecular forms reacted to all 3 CCK-antibodies it is likely that they contain the sulfated tyrosyl and carboxy-terminal regions of CCK and, therefore, possess biological activity.