The isolation, purification and some properties of pheromaxein, the pheromonal steroid-binding protein, in porcine submaxillary glands and saliva.
Pheromaxein, the 16-androstene steroid-binding protein with a relative molecular mass of 15,000 was isolated in sub-milligram quantities from the submaxillary gland and saliva of the Gottingen miniature boar, after a fourfold purification involving the following methods: ultrafiltration for submaxillary gland cytosols and ethanol precipitation for saliva, Concanavalin-A-Sepharose affinity chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis, 'Extractigel-D' affinity chromatography (to remove sodium dodecyl sulphate) and fast protein-liquid chromatography. Yields of purified pheromaxein obtained after fast protein-liquid chromatography represented 10-20% of total protein present in an ultrafiltrate of a submaxillary gland cytosol. Fast protein-liquid chromatography separated the alpha- and beta-charge isomers of pheromaxein which were shown to have isoelectric points of 4.78 and 5.35 respectively on flat-bed isoelectric focusing. Some data are provided for the variable occurrence of the isomeric forms of pheromaxein in relation to different breeds of pig. Five 16-unsaturated steroids showed the highest binding to pheromaxein. Other steroids of the 5 alpha- and 5 beta-androstane series also showed some binding to pheromaxein, i.e. 17 beta-hydroxy-5 alpha-androstan-3-one (19.2%), with 5 alpha-androstan-3-one, which has a similar urinous odour to 5 alpha-androst-16-en-3-one, showing the greatest binding (42.6%) relative to 5 alpha-androst-16-en-3-one (100%).
Booth
W D
WD
AFRC Institute of Animal Physiology and Genetics Research, Cambridge.
White
C A
CA
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D012463:Saliva / Q000032:analysis*; D013363:Submandibular Gland / Q000032:analysis*; D013552:Swine; D013556:Swine, Miniature / Q000378:metabolism*
D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; C054734:pheromaxein