Cell-free synthesis, proteolytic processing, core glycosylation, and amino terminal sequence of rabbit pre-alpha-lactalbumin.
Two different forms of alpha-lactalbumin were isolated from rabbit milk and partially characterized. The major and the minor species had apparent molecular weights of 18000 and 14000, respectively, according to their electrophoretic mobilities on SDS polyacrylamide gels. Analyses of their amino acid compositions and amino-and carboxy-terminal sequences did not reveal any difference, but sugar analysis showed the occurrence of carbohydrates in the major species. Rabbit alpha-lactalbumin was synthesized in a cell-free translation system as a precursor with an amino terminal extension of 19 amino acid residues whose primary structure is rather different from those of its ovine and porcine counterparts, in contrast with the extensive similarity so far observed between the known signals of homologous milk proteins. When mammary microsomal membranes were added during translation, the preprotein was converted to authentic alpha-lactalbumin, as demonstrated by amino terminal sequence analyses. However, one of the two processed forms migrated more slowly than pre-alpha-lactalbumin on SDS polyacrylamide gels and this was related to the occurrence of carbohydrates: only the "slower moving" polypeptide was specifically adsorbed on concanavalin A Sepharose and its electrophoretic mobility was enhanced after treatment with endoglycosidase H, an enzyme known to remove clustered mannosyl residues linked to di-N-acetylchitobiose. It was also observed that the rate of translocation of alpha-lactalbumin across the microsomal membrane was lower than that of beta-casein.
D000595:Amino Acid Sequence; D000818:Animals; D050260:Carbohydrate Metabolism; D002474:Cell-Free System; D005260:Female; D007425:Intracellular Membranes / Q000378:metabolism; D007768:Lactalbumin / Q000096:biosynthesis / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D008892:Milk / Q000378:metabolism*; D008970:Molecular Weight; D011247:Pregnancy; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism; D011817:Rabbits
D011498:Protein Precursors; C034435:pre-alpha-lactalbumin; D007768:Lactalbumin