The porcine trophoblastic interferon-gamma, secreted by a polarized epithelium, has specific structural and biochemical properties.
At the time of implantation in the maternal uterus, the trophectoderm of the pig blastocyst is the source of a massive secretion of interferon-gamma (IFN-gamma), together with lesser amounts of IFN-delta, a unique species of type I IFN. This trophoblastic IFN-gamma (TrIFN-gamma) is an unprecedented example of IFN-gamma being produced spontaneously by an epithelium. We therefore studied some of its structural and biochemical properties, by comparison with pig IFN-gamma from other sources, either natural LeIFN-gamma (from adult leucocytes), or recombinant. Biologically active TrIFN-gamma is a dimeric molecule, of which monomers are mainly composed of a truncated polypeptide chain with two glycotypes, unlike LeIFN-gamma which is formed of at least two polypeptide chains and four glycotypes. TrIFN-gamma collected in the uterus lumen was enzymatically deglycosylated and analysed by mass spectrometry (MALDI-TOF). The data revealed that the more abundant polypeptide has a mass of 14.74 kDa, corresponding to a C-terminal cleavage of 17 residues from the expected 143-residue long mature sequence. A minor polypeptide, with a mass of 12.63 kDa, corresponds to a C-terminal truncation of 36 amino acids. MALDI-TOF analysis of tryptic peptides from the glycosylated molecule(s) identifies a single branched carbohydrate motif, with six N-acetylgalactosamines, and no sialic acid. The only glycan microheterogeneity seems to reside in the number of l-fucose residues (one to three). The lack of the C-terminal cluster of basic residues, and the presence of nonsialylated glycans, result in a very low net charge of TrIFN-gamma molecule. However, the 17-residue truncation does not affect the antiproliferative activity of TrIFN-gamma on different cells, among which is a porcine uterine epithelial cell line. It is suggested that these specific properties might confer on TrIFN-gamma a particular ability to invade the uterine mucosa and exert biological functions beyond the endometrial epithelium.
Cencic
Avrelija
A
Unité de Virologie et d'Immunologie Moléculaires, INRA, Jouy en Josas, France.
Henry
Céline
C
Lefèvre
François
F
Huet
Jean-Claude
JC
Koren
Srecko
S
La Bonnardière
Claude
C
D000818:Animals; D016764:Cell Polarity; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D007371:Interferon-gamma / Q000737:chemistry / Q000378:metabolism*; D007962:Leukocytes / Q000378:metabolism; D013552:Swine; D014327:Trophoblasts / Q000378:metabolism*
D007371:Interferon-gamma