Related Experiment Videos
Characterization of proliferin-related protein
P Colosi1, J J Swiergiel, E L Wilder
1Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208.
Abstract:
Proliferin-related protein (mPRP) is a member of the PRL/GH family in the mouse. We have generated an antiserum against mPRP expressed as a bacterial fusion protein; this antiserum detects mPRP in the conditioned media of placental tissue cultures as a heterogeneous population of glycoproteins. We have also expressed mPRP in mammalian tissue culture cells and purified the secreted protein. N-terminal sequence analysis of the purified protein reveals that it is secreted as a 214 amino acid protein after removal of a 30 amino acid signal polypeptide. An antiserum raised against the purified protein detects high levels of mPRP in maternal serum during gestation. The site of synthesis of this protein has been localized by in situ hybridization to the basal zone of the day-10 mouse placenta, which is distinct from the site of synthesis of other placental proteins in this family.
Insights
Researchers developed an antiserum to detect mouse prolifern-related protein (mPRP), a placental glycoprotein. High levels of mPRP were found in maternal serum during mouse gestation.
Area of Science:
- Reproductive biology
- Molecular endocrinology
- Protein biochemistry
Background:
- Proliferin-related protein (mPRP) is part of the prolactin/growth hormone (PRL/GH) family in mice.
- Understanding mPRP's role requires characterizing its expression and localization.
Purpose of the Study:
- To generate and characterize an antiserum for mouse prolifern-related protein (mPRP).
- To investigate the secretion, purification, and localization of mPRP during mouse gestation.
Main Methods:
- Bacterial fusion protein expression and antiserum generation.
- Mammalian cell culture for mPRP expression and purification.
- N-terminal sequencing for protein characterization.
- In situ hybridization for placental localization.
Main Results:
- An antiserum detected mPRP as heterogeneous glycoproteins in placental conditioned media.
- Purified mPRP is secreted as a 214-amino acid protein after signal peptide cleavage.
- High mPRP levels were observed in maternal serum during gestation.
- mPRP synthesis was localized to the basal zone of the day-10 mouse placenta.
Conclusions:
- The study successfully generated specific antibodies against mPRP.
- mPRP is a secreted glycoprotein found in maternal serum during pregnancy.
- The unique placental localization suggests a specific role for mPRP in mouse gestation.