Growth hormone heterogeneity in American plaice pituitaries: isolation, characterization, and partial amino acid sequence.

Growth hormones (GHs) have been isolated from pituitary glands of American plaice (Hippoglossoides platessoides), a marine flatfish, using affinity and gel filtration chromatography, followed by preparative polyacrylamide gel electrophoresis (PAGE). A bioassay based on serum triiodothyronine elevation in immature rainbow trout was used to monitor biological activity. These GHs originate from two molecular mass regions, 42K and less than 33K relative molecular mass (Mr), in their native state. The 42K Mr region yielded two forms of GH, which differ in terms of quantity and net charge as evidenced by native PAGE, a major variant with a relative mobility of (Rf) 0.22 and a lesser variant with Rf 0.28. The less than 33 Mr region has a single GH species with Rf 0.22. Upon sodium dodecyl sulfate-PAGE, without reduction, both GH variants from the 42K Mr region gave Mrs of 21K, while the GH from the less than 33K Mr region was 20K Mr, typical of monomeric vertebrate GHs. The proteins composing the 42K Mr region are proposed as GH dimers since they yield 21K Mr peptides. The less than 33K Mr region contains a GH monomer (20K Mr) in its native state. An amino-terminal amino acid sequence, identical for both the 42K and the 20K Mr Rf 0.22 forms, has good homology with other complete fish GH sequences near their carboxyl-terminal regions (between amino acids 130 and 196). The GH dimers (42K Mr) predominate in the plaice pituitary, contributing 93% of the total, of which 86% gives rise to the Rf 0.22 variant.