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Studies of the biosynthesis of renin with a cell-free translation system
Abstract:
1. Poly(A)+ mRNA from mouse submaxillary gland encodes a polypeptide of molecular weight 48 000 (48K polypeptide) which is abundant in the male. 2. This polypeptide is selectively absent in the translation products of mRNA from a strain of genetically renin-deficient mice C57 BL/10J. 3. The 48K polypeptide binds and co-elutes in identical fashion with pure authentic renin on pepstatin affinity chromatography. 4. Immunoprecipitation of translation products of male glandular mRNA with renin-specific antibody yielded this 48K band upon analysis by SDS/polyacrylamide gel electrophoresis and fluorography. Pure renin of molecular weight 37 000 blocked the binding of this polypeptide to antirenin antibody. 5. Mouse submaxillary gland synthesizes a renin precursor. The renin mRNA is androgenically regulated.
Insights
Mouse submaxillary glands produce a 48K polypeptide abundant in males, identified as a renin precursor. This finding reveals androgenic regulation of renin mRNA in mice.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- The mouse submaxillary gland is known to express various proteins, including enzymes like renin.
- Previous studies have indicated sexual dimorphism in the expression of certain proteins in this gland.
Purpose of the Study:
- To identify and characterize a specific polypeptide encoded by polyadenylated mRNA from the male mouse submaxillary gland.
- To determine if this polypeptide is related to renin and investigate its regulation.
Main Methods:
- Poly(A)+ mRNA isolation and in vitro translation.
- Pepstatin affinity chromatography for protein binding analysis.
- Immunoprecipitation using renin-specific antibodies.
- SDS/polyacrylamide gel electrophoresis and fluorography for molecular weight analysis.
Main Results:
- A 48,000 molecular weight (48K) polypeptide was identified, abundant in male mouse submaxillary gland mRNA translation products.
- This 48K polypeptide was absent in renin-deficient mice (C57 BL/10J).
- The 48K polypeptide exhibited identical binding and co-elution with authentic renin on pepstatin affinity chromatography and was immunoprecipitated by renin-specific antibodies, confirming its identity as a renin precursor. Pure renin (37,000 MW) blocked this binding.
- Renin mRNA was found to be androgenically regulated.
Conclusions:
- The mouse submaxillary gland synthesizes a renin precursor polypeptide.
- The expression of renin mRNA in this gland is regulated by androgens.