Several intermediate forms in the processing of rat lutropin subunits as shown by sodium dodecyl sulphate polyacrylamide gel electrophoresis.
To study the biosynthesis in situ of lutropin (LH) subunits, anterior pituitary cells in culture were employed. The cells were incubated in the presence of [35S]methionine. Labelled polypeptides, immunologically related to alpha and LH beta subunits, were isolated by specific immunoprecipitation, then analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and revealed by fluorography. Electrophoretic analysis of immunoprecipitates is a crucial step in this methodology. It permits simultaneous purification, characterization and accurate quantitation of the radioactivity specifically incorporated into LH subunits. Also, using SDS-PAGE, it is possible to isolate and identify the different processed forms of LH subunits.
Starzec
A
A
Laboratoire des Hormones Polypeptidiques, CNRS, Gif-sur-Yvette, France.
Counis
R
R
Jutisz
M
M
D000818:Animals; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D007120:Immunochemistry; D007986:Luteinizing Hormone / Q000096:biosynthesis / Q000302:isolation & purification*; D008297:Male; D010903:Pituitary Gland, Anterior / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012967:Sodium Dodecyl Sulfate; D014415:Tunicamycin / Q000302:isolation & purification
D014415:Tunicamycin; D012967:Sodium Dodecyl Sulfate; D007986:Luteinizing Hormone