Biological, immunological, and binding properties of recombinant mouse placental lactogen-I

Mouse placental lactogen-I (mPL-I) cDNA was inserted into a eukaryotic expression vector and introduced into Chinese hamster ovary cells. Cell lines that secrete high concentrations of mPL-I were isolated, and this glycoprotein was purified from the cell culture-conditioned medium. Recombinant mPL-I...

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Published inEndocrinology (Philadelphia) Vol. 123; no. 6; p. 2662
Main Authors Colosi, P, Ogren, L, Southard, J N, Thordarson, G, Linzer, D I, Talamantes, F
Format Journal Article
LanguageEnglish
Published United States 01.12.1988
Subjects
Animals
Biological Assay
Cell Line
Cricetinae
DNA - genetics
Electrophoresis, Polyacrylamide Gel
Epithelium - metabolism
Lactalbumin - biosynthesis
Macromolecular Substances
Mammary Glands, Animal - drug effects
Mammary Glands, Animal - metabolism
Mice
Microsomes, Liver - metabolism
Molecular Weight
Placental Lactogen - genetics
Placental Lactogen - metabolism
Placental Lactogen - pharmacology
Radioimmunoassay
Radioligand Assay
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Recombinant Proteins - pharmacology
Transfection
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Summary:Mouse placental lactogen-I (mPL-I) cDNA was inserted into a eukaryotic expression vector and introduced into Chinese hamster ovary cells. Cell lines that secrete high concentrations of mPL-I were isolated, and this glycoprotein was purified from the cell culture-conditioned medium. Recombinant mPL-I (mPL-Ir) is very similar to placental mPL-I (mPL-Ip) in its recognition by polyclonal antisera raised against either mPL-Ip or mPL-Ir, in displacing [125I]iodo-mPL-II from binding sites on mouse liver microsomal membranes, and in stimulating the synthesis of alpha-lactalbumin in primary cultures of mouse mammary epithelial cells. Structural comparison of mPL-Ir and mPL-Ip by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that mPL-Ir comprises several proteins with mol wt ranging from 34.5-38K, while mPL-Ip consists of a similar set of proteins with mol wt ranging from 36.5-42K. Treatment of the two proteins with neuraminidase resulted in similar 2-4K decreases in mol wt. Treatment of mPL-Ip with peptide:N-glycosidase-F to remove asparagine-linked oligosaccharide chains resulted in the formation of 28K and 29K mol wt species, while treatment of mPL-Ir with the same enzyme yielded 28K and 28.5K mol wt products.
ISSN:0013-7227
DOI:10.1210/endo-123-6-2662