Characterization of feline conceptus proteins during pregnancy.
This study characterized proteins secreted de novo by feline conceptuses collected on Days 10, 12, and 15 (n = 22, preimplantation blastocysts); Days 15, 16, 17, 19, 21, and 25 (n = 6, postimplantation zonary girdle [ZG] i.e. trophoblast and endometrium); and Days 30, 36, 39, and 50 (n = 5, postimplantation ZG and free chorioallantois [CA]) and cultured in Minimal Essential Medium. De novo secretion was shown by incorporation of 3H-leucine into proteins detected in culture media by 2D-PAGE and fluorography. Western blotting, and NH2-terminal amino acid microsequencing. Major radiolabeled proteins identified as they appeared temporally on fluorographs were as follows: feline conceptus protein 1 (fCP1), Mr = 20,000, pI 5.0-5.3; fCP2, Mr = 80,000, pI 6.5-7.2; fCP3a, Mr = 67,000, pI 6.3-6.5; fCP3b, Mr = 67,000, pI 5.9-6.3; fCP4, Mr = 56,000, pI 5.0-6.0; and fCP5, Mr = 29,000, pI 5.0-5.8. The fCP1 was produced by blastocysts on Days 10-15, ZG on Days 16-25, and CA on Day 30; on Days 39-50, CA synthesized 5 proteins, possibly fCP1 isomers. The fCP2, fCP3a and b, and fCP4 were produced by blastocysts on Day 15, ZG on Day 25, and CA on Days 30-50. The fCP5 was made by ZG on Days 16-36 and by CA on Days 30-39. Western blotting identified fCP1 as retinol-binding protein (RBP), fCP2 as alpha fetoprotein, fCP3a as albumin, and fCP3b as transferrin. Amino acid sequence homologies between fCP1 and rabbit and human plasma RBP and porcine conceptus RBP2 were 93, 96, and 100%, respectively, at the first 37 NH2-terminal amino acids. The identities of fCP4 and fCP5 have not been established. Antiviral activity detected in all media was less than 3 units/ml when tested with feline fibroblast cells infected with vesicular stomatitis virus.
Thatcher
M D
MD
College of Veterinary Medicine, University of Florida, Gainesville 32610.
Shille
V M
VM
Fliss
M F
MF
Bazer
F W
FW
Sisum
W
W
Randal
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002415:Cats; D004622:Embryo, Mammalian / Q000378:metabolism; D004717:Endometrium / Q000378:metabolism; D005321:Extraembryonic Membranes / Q000378:metabolism; D005260:Female; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D054839:Retinol-Binding Proteins, Plasma; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors
D011257:Pregnancy Proteins; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma