Mammary Derived Growth Inhibitor Is Not a Distinct Protein but a Mix of Heart-type and Adipocyte-type Fatty Acid-binding Protein
| Autor: | Norbert Bartetzko, Helena Kuhl, Bernfried Specht, Carsten Hohoff, Friedrich Spener, Torsten Börchers, Regina Franke |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Immunoprecipitation
Blotting Western Molecular Sequence Data Gene Expression Nerve Tissue Proteins Biology Fatty Acid-Binding Proteins Myelin P2 Protein Biochemistry law.invention Mice Mammary Glands Animal law Complementary DNA Animals Humans Electrophoresis Gel Two-Dimensional Amino Acid Sequence Isoelectric Point RNA Messenger Fluorescent Antibody Technique Indirect Molecular Biology Peptide sequence DNA Primers Messenger RNA Base Sequence Sequence Homology Amino Acid Isoelectric focusing Myocardium Tumor Suppressor Proteins Cell Biology Molecular biology In vitro Neoplasm Proteins Blot Adipose Tissue Recombinant DNA Cattle Female lipids (amino acids peptides and proteins) Carrier Proteins Fatty Acid-Binding Protein 7 Fatty Acid Binding Protein 3 Sequence Alignment |
| Zdroj: | Journal of Biological Chemistry. 271:19943-19949 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.271.33.19943 |
| Popis: | The amino acid sequence of the mammary derived growth inhibitor (MDGI) from bovine mammary gland (Böhmer, F.-D., Kraft, R., Otto, A. , Wernstedt, C., Hellman, U., Kurtz, A., Müller, T., Rohde, K., Etzold, G., Lehmann, W., Langen, P., Heldin, C.-H., and Grosse, R. (1987) J. Biol. Chem. 262, 15137-15143) revealed 95% identity to bovine heart fatty acid-binding protein (H-FABP), explaining the observed immunocross-reactivity. However, a cDNA encoding MDGI has not been found to date. Artificial MDGI cDNA was expressed in an in vitro transcription/translation assay. Analysis by isoelectric focusing of the immunoprecipitated in vitro translation products of lactating bovine mammary gland mRNA did not indicate a protein corresponding to the in vitro translation product of artificial MDGI mRNA. Moreover, two-dimensional electrophoresis of bovine mammary gland proteins confirmed the absence of a protein with the pI of the in vitro translated artificial MDGI mRNA in bovine mammary gland and instead revealed, apart from H-FABP, an unknown protein that was recognized by anti-H-FABP antibodies. From lactating bovine mammary gland the cDNA for adipocyte fatty acid-binding protein (A-FABP) was cloned. The in vitro translation of recombinant mRNA derived from this cDNA yielded a polypeptide that behaved like the unknown immunoreactive protein. Western blotting and immunofluorescence using monospecific antibodies demonstrated the coexistence of H-FABP and A-FABP in the lactating mammary gland. Taking into account that deviations of the MDGI sequence from the bovine H-FABP sequence correspond with A-FABP we attribute the structure originally reported as MDGI to a mix of these proteins. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|