Identification of multiple pregnancy-associated glycoproteins (PAGs) purified from the European bison (Eb; Bison bonasus L.) placentas
Kiewisz, J. | Sousa, N Melo de | Beckers, J.F. | Panasiewicz, G. | Gizejewski, Z. | Szafranska, B.
This paper describes the first identified chorionic PAGs in the European bison (Eb), named EbPAGs, predominantly expressed during early and mid-pregnancy (45-120 day post-coitum; dpc). Many EbPAGs were extracted from various cotyledonary tissues, precipitated, chromatographed (DEAE and VVA: Vicia villosa agglutinin), electrophoresed (1D- and 2D-PAGE), analysed by heterologous (cross-species) Western blotting and then micro-sequenced by Edman degradation. Finally, twelve selected VVA-purified isoforms (Ip 3.7-7.4) were entirely characterised. Nine identified NH₂-terminal micro-sequences were found to be PAGs. On 45dpc, three identified forms were named: EbPAG₆₇A kDa (RGSNLTHPLRNIGDLFYVGN), EbPAG₅₅B kDa (RGSNLTHPL) and EbPAG₅₀C kDa (SQISLRGSNLTI). On 60dpc, the next three forms were named: EbPAG₇₁D kDa (RGSNLTIHPLRNIIDLFYVG), EbPAG₅₅E kDa (RGSNLTHPLRNI) and EbPAG₅₀F kDa (SQISLRGS). On 120dpc, three other forms were named: EbPAG₇₁G kDa (RGSNLTHPLRNIRDLFYVG), EbPAG₆₀H kDa (RGSNLTTHPLRNIKDLVVYM) and EbPAG₅₀I kDa (SGSNLTTV). These EbPAG (A₋I) sequences are unique, as they are not identical to any other PAGs purified previously in related species of the Bovidae family. However, the EbPAGs (A-I forms) have some sequence resemblance to internal sequences of various full-length polypeptide PAG precursors (in silico translated from cloned cDNAs) identified in domestic cattle. Three other novel native isoforms (J1, J2 and K): EbUPG₄₅kDa J1 (SKDNYKNYIPLIVPFAT), EbUPG₄₅kDa J2 (SKDNQKNYIPLIVPFAT) and EbUPG₇₆kDa K (SPEFTV), were temporarily named 'unknown placental glycoproteins' (UPGs), due to their efficient VVA-purification (specific for glycoproteins only) and a lack of considerable consensus to previously sequenced placental glycoproteins in the Bovidae family. This is the first study identifying NH₂-terminals of multiple/diverse EbPAGs and some EbUPGs purified from the synepitheliochorial cotyledonary placenta of the endangered Bison bonasus (Red List).
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