Isolation and characterization of three forms of luteinizing hormone from the pituitary gland of the horse.

Three isoforms of equine luteinizing hormone (eLH-A, eLH-B and eLH-C) have been isolated from horse pituitary glands. Separation was achieved on the basis of charge heterogeneity by ion-exchange chromatography. These charge differences were apparent after final purification, as determined by electrophoretic mobility on polyacrylamide disc gels (RF = 0.14, 0.19 and 0.26 for eLH-A, -B and -C, respectively). Apparent size differences were also noted between the isohormones by gel filtration on Sephadex G-100. Ve/Vo ratios for eLH-A, -B and -C were 1.72, 1.54 and 1.47, respectively. All 3 isoforms were found to contain an equivalent amount of hexose (9.0-9.2%). Isohormones eLH-B and eLH-C, however, possess more sialic acid than eLH-A (6.6-6.7%, vs. 4.5%). The eLH-A and eLH-B preparations contain a similar amount of hexosamine, which is slightly lower than the amount of eLH-C (8.8-9.1% vs. 11.2%). No differences were noted between the isohormones by rat Leydig cell LH bioassay, equine testis LH radioreceptor assay (RRA) or calf testis follicle-stimulating hormone (FSH) RRA. Slight, but nonsignificant, variations were noted between preparations in an eLH radioimmunoassay (RIA). Although chemical variations were detected between the eLH isoforms, no significant differences were observed in in vitro biological and immunological activities. The differences detected in sialic acid content raises the possibility that differences in in vivo clearance rates may exist.