Placental lactogen-I (PL-I) target tissues identified with an alkaline phosphatase-PL-I fusion protein.
The rat placenta expresses a family of genes related to prolactin (PRL). Target tissues and physiological roles for many members of the PRL family have yet to be determined. In this investigation we evaluated the use of an alkaline phosphatase (AP) tag for monitoring the behavior of a prototypical member of the PRL family, placental lactogen-I (PL-I). A probe was generated consisting of a fusion protein of human placental AP and rat PL-I (AP-PL-I). The AP-PL-I construct was stably expressed in 293 human fetal kidney cells, as was the unmodified AP vector that served as a control. AP activity was monitored with a colorimetric assay in conditioned medium from transfected cells. Immunoreactivity and PRL-like biological activities of the AP-PL-I fusion protein were demonstrated by immunoblotting and the Nb2 lymphoma cell proliferation assay, respectively. AP-PL-I specifically bound to tissue sections known to express the PRL receptor, including the ovary, liver, and choroid plexus. Binding of AP-PL-I to tissues was specific and could be competed with ovine PRL. The results indicate that AP is an effective tag for monitoring the behavior of PL-I and suggest that this labeling system may also be useful for monitoring the actions of other members of the PRL family.
Müller
H
H
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160-7401, USA.
Dai
G
G
Soares
M J
MJ
D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D015153:Blotting, Western; D002831:Choroid Plexus / Q000737:chemistry; D005260:Female; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000737:chemistry; D008223:Lymphoma / Q000737:chemistry; D010053:Ovary / Q000737:chemistry; D010928:Placental Lactogen / Q000032:analysis* / Q000276:immunology; D011247:Pregnancy; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000032:analysis; D014407:Tumor Cells, Cultured
D011993:Recombinant Fusion Proteins; C085699:placental lactogen I, rat; D010928:Placental Lactogen; D000469:Alkaline Phosphatase