Bovine prolactin-related protein-I is anchored to the extracellular matrix through interactions with type IV collagen.
The bovine placenta produces an array of proteins structurally similar to pituitary prolactin (PRL). At least ten genes of the bovine placental PRL family, including bovine placental lactogen (bPL) and ten bovine PRL-related protein-I to -X (bPRP-I to -X), encode hormones/cytokines predicted to be involved in the establishment and maintenance of pregnancy. Targets and biological roles for most members of the bovine PRL family have yet to be specified. This study focused on three members of bovine PRL family, bPL, bPRP-I, and bPRP-VI. An alkaline phosphatase (AP) tagging strategy was used to monitor interactions of the ligands with their targets. AP-bPRP-I and AP-bPRP-VI specifically bound to tissue sections of the bovine placentome. AP-bPRP-I and AP-bPRP-VI binding within the placentome mimicked the distribution of the extracellular matrix (ECM). Consequently, AP fusion protein binding to individual ECM components (heparin, laminin, fibronectin, type I collagen, and type IV collagen) was evaluated. AP-bPRP-I specifically bound to type IV collagen, but not to the other ECM components. AP-bPRP-VI exhibited weak interactions with ECM components, while AP-bPL and AP did not show significant binding to any of the ECM components. Binding of AP-bPRP-I to type IV collagen was concentration-dependent, influenced by salt concentrations, and specific to the N-terminal cross-linking domain (7S) of type IV collagen but not its triple-helical domain. The interaction of bPRP-I with type IV collagen suggests that bPRP-I accumulates in the ECM where it likely acts on cells traversing the bovine placentome.
Takahashi
Toru
T
Reproductive Biology Research Unit, National Institute of Agrobiological Sciences, Tsukuba City, Ibaraki 305-8602, Japan.
Yamada
Osamu
O
Soares
Michael J
MJ
Hashizume
Kazuyoshi
K
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D002417:Cattle; D002460:Cell Line; D024141:Collagen Type IV / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005260:Female; D006801:Humans; D009994:Osmolar Concentration; D010920:Placenta / Q000378:metabolism*; D011388:Prolactin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014018:Tissue Distribution
D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D011993:Recombinant Fusion Proteins; D011388:Prolactin; D000469:Alkaline Phosphatase