Prosaposin, a neurotrophic factor: presence and properties in milk.
The presence of prosaposin, the precursor of the sphingolipid activator proteins (saposins A, B, C, and D), was investigated in bovine milk. The milk proteins were resolved by SDS-PAGE, blotted onto nitrocellulose sheets, and immunostained. Each of three appropriate antibodies defined a band from milk that matched in mobility the reference prosaposin from human milk at a relative molecular mass of 66,000. Evidence of mature saposins was not found. Prosaposin was detected in milk of other species chimpanzee, rhesus, goat, and rat) and was consistently observed in samples of retail milk and from individual cows. Prosaposin was not associated with particulate matter (fat globules, casein micelles, membrane fragments, and somatic cells) in either human or bovine milk. Rather, prosaposin was located exclusively in the milk serum (whey), existing in monomeric form, as revealed by nondenaturing PAGE. A commercial whey protein concentrate (75% protein) appeared to retain milk prosaposin quantitatively. Properties that were useful in the isolation of prosaposin from milk were its binding to concanavalin A, retention by anion-exchange cellulose, and resistance to precipitation by heating. The possibility that bovine milk prosaposin nutritionally benefits the humans who consume it is enhanced by the fact that only part of its saposin C segment is required for neurotrophic activity.
Patton
S
S
Department of Neurosciences, University of California San Diego, La Jolla 92093, USA.
Carson
G S
GS
Hiraiwa
M
M
O'Brien
J S
JS
Sano
A
A
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D003208:Concanavalin A / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008253:Macaca mulatta; D008892:Milk / Q000737:chemistry*; D008895:Milk, Human / Q000737:chemistry; D008970:Molecular Weight; D002679:Pan troglodytes; D051381:Rats; D049231:Saposins
D006023:Glycoproteins; C483345:PSAP protein, human; C483347:Psap protein, rat; D049231:Saposins; D003208:Concanavalin A