The absence from the oocyte secretory apparatus of a protein kinase capable of phosphorylating sequestered caseins.
The lactating guinea-pig mammary gland synthesizes and secretes four major milk proteins, i.e., three caseins and alpha-lactalbumin. Of these, the caseins are highly phosphorylated, a post-translational event which in the mammary gland involves a specific casein kinase, which is an integral membrane protein probably of Golgi origin. The microinjection of milk protein mRNA into Xenopus oocytes in the presence of [35S]methionine leads to the synthesis, sequestration, and secretion of proteins which coelectrophorese with alpha-lactalbumin and with partially processed caseins. That the secreted caseins were not phosphorylated was shown by the use of 32P. Either the oocytes were injected with mammary gland mRNA followed by incubation with [32P]phosphate containing media or the mRNA was co-injected with [gamma-32P]ATP and the oocytes were then incubated. In neither case were 32P-labeled caseins secreted. Golgi-rich fractions, identified by the marker enzyme galactosyltransferase, were isolated from the postnuclear supernatant of both oocytes and lactating mammary gland by sucrose density gradient fractionation. In contrast to the mammary gland fractions those derived from the oocytes contained no detectable casein kinase activity. Homogenates of oocytes do effect the phosphorylation of casein but the enzyme activity appears to be present in the soluble fraction and is not membrane bound. It is concluded that the Xenopus oocyte lacks the specific kinase that in the mammary gland phosphorylates sequestered caseins and that the phosphorylation of the caseins is not a prerequisite for their secretion by the oocyte.
A P
AP
Boulton
C D
CD
Lane
J C
JC
Pascall
R K
RK
Craig
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000378:metabolism*; D002474:Cell-Free System; D005260:Female; D006168:Guinea Pigs; D007774:Lactation; D008321:Mammary Glands, Animal / Q000378:metabolism; D009865:Oocytes / Q000201:enzymology*; D010766:Phosphorylation; D011061:Poly A / Q000378:metabolism; D011247:Pregnancy; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D014908:Triticum; D014981:Xenopus
D002364:Caseins; D011061:Poly A; D012313:RNA; D000255:Adenosine Triphosphate; D011494:Protein Kinases