Purification, characterization, and immunocytochemical localization of the major basic protein of pig blastocysts.
The major basic protein (BP) synthesized and secreted by elongating pig blastocysts was purified from medium of Day 14-17 conceptus cultures. Sequential ion-exchange and gel-filtration chromatographies resulted in isolation of BP as a single polypeptide of Mr = 43,100 or 42,800 under denaturing or native conditions, respectively. BP was found to be a glycoprotein by incorporation of [3H] glucosamine and susceptibility to N-glycopeptidase F. Two BP polypeptides were produced by N-glycopeptidase F (Mr = 39,800 and 36,300). Antiserum to BP immunoprecipitated radiolabeled BP from blastocyst culture medium. BP was not detected in medium from 1-2 mm diameter spherical (Day 10) blastocysts but was found in medium from 3-5 mm spherical (Day 10) and filamentous (less than 50 cm, Day 12) conceptuses, suggesting that BP synthesis and secretion began at the initiation of trophoblast expansion. With immunocytochemical procedures, BP was located in the apical cytoplasm of trophectoderm cells of Day 11 expanding (5-7 and 10-20 mm) blastocysts. These results suggest that trophoblast epithelium secrete BP apically toward the uterine lumen and that BP may play a role in maternal-fetal interactions during the peri-implantation period.
Baumbach
G A
GA
Department of Animal Science, College of Veterinary Medicine, University of Tennessee, Knoxville 37901.
Climer
A H
AH
Bartley
N G
NG
Kattesh
H G
HG
Godkin
J D
JD
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D001798:Blood Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D002478:Cells, Cultured; D047091:Eosinophil Granule Proteins; D005260:Female; D010452:Peptide Biosynthesis; D010455:Peptides / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D012260:Ribonucleases; D013552:Swine / Q000378:metabolism*
D001798:Blood Proteins; D047091:Eosinophil Granule Proteins; D010455:Peptides; D012260:Ribonucleases