Hostname: page-component-586b7cd67f-r5fsc Total loading time: 0 Render date: 2024-11-23T11:25:19.375Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and in vitro translation of mRNA from the calf abomasal mucosa and identification of an mRNA coding for a precursor of prochymosin

Published online by Cambridge University Press:  01 June 2009

Bruce H. Nicholson  and
Peter Jones
Bruce H. Nicholson
Affiliation:
Department of Physiology and Biochemistry, The University, Whiteknights, Reading RG6 2AJ, UK
Peter Jones
Affiliation:
Department of Physiology and Biochemistry, The University, Whiteknights, Reading RG6 2AJ, UK
Get access Rights & Permissions [Opens in a new window]

Summary

The mRNA coding for prochymosin (prorennin) has been partly purified from calf abomasum. The in vitro translation products of the total polyadenylated RNA show a major band on gel electrophoresis which reacts with antibody raised against purified chymosin. The mol. wt of 43000 is higher than expected from the reported amino acid sequence and would correspond to prochymosin with an unprocessed signal sequence of ∼ 17 amino acids. The synthesis of chymosin mRNA is age-related, and ceases by 3 months even in milk-fed calves.

Type
Original Articles
Information
Journal of Dairy Research , Volume 51 , Issue 1 , February 1984 , pp. 79 - 89
Copyright
Copyright © Proprietors of Journal of Dairy Research 1984

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Blobel, G. & Dobberstein, B. 1975 a Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. Journal of Cell Biology 67 835–851CrossRefGoogle ScholarPubMed
Blobel, G. & Dobberstein, B. 1975 b Transfer of proteins across membranes. II. Reconstitution of functional rough microsomes from heterologous components. Journal of Cell Biology 67 852–862CrossRefGoogle ScholarPubMed
Blundell, T. L., Jones, H. B., Khan, G., Taylor, G., Sewell, B. T., Pearl, L. H. & Wood, S. P. 1980 The active site of acid proteinases. In Enzyme regulation and mechanism of action, pp. 281–288. (Eds Mildner, P. and Ries, B.). Oxford: Pergamon PressCrossRefGoogle Scholar
Bonner, W. M. & Laskey, R. A. 1974 A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46 83–88CrossRefGoogle ScholarPubMed
Craig, R. K., Perera, P. A. J., Mellor, A. & Smith, A. E. 1979 Initiation and processing in vitro of the primary translation products of guinea-pig caseins. Biochemical Journal 184 261–267CrossRefGoogle ScholarPubMed
Foltmann, B. 1970 Prochymosin and chymosin (prorennin and rennin). Methods in Enzymology 19 421–436CrossRefGoogle Scholar
Foltmann, B. 1981 Gastric proteinases – structure, function, evolution and mechanism of action. In Essays in Biochemistry 17, pp. 52–84 (Eds Campbell, P. N. and Marshall, R. D.). London. Academic PressGoogle Scholar
Foltmann, B., Jensen, A. L., Lonblad, P., Smidt, E. & Axelsen, N. H. 1981 A developmental analysis of the production of chymosin and pepsin in pigs. Comparative Biochemistry and Physiology 68 B 9–13Google Scholar
Foltmann, B., Pedersen, V. B., Jacobsen, H., Kauffman, D. & Wybrandt, G. 1977 The complete amino acid sequence of prochymosin. Proceedings of the National Academy of Sciences of the USA 74 2321–2324CrossRefGoogle ScholarPubMed
Foltmann, B., Pedersen, V. B., Kauffman, D. & Wybrandt, G. 1979 The primary structure of calf chymosin. Journal of Biological Chemistry 254 8447–8456CrossRefGoogle ScholarPubMed
Garnot, P., Toulleo, R., Thapon, J. L., Martin, P., Hoano, M. T., Mathieu, C. M. & Ribadeau Dumas, B. 1977 Influence of age, dietary protein and weaning on calf abomasal enzymic secretion. Journal of Dairy Research 44 9–23CrossRefGoogle ScholarPubMed
Harris, T. J. R., Lowe, P. A., Lyons, A., Thomas, P. G., Eaton, M. A. W., Millican, T. A., Patel, T. P., Bose, C. C., Carey, N. H. & Doet, M. T. 1982 Molecular cloning and nucleotide sequence of cDNA coding for calf preprochymosin. Nucleic Acids Research 10 2177–2187CrossRefGoogle ScholarPubMed
Hensohel, M. J., Hill, W. B. & Porter, J. W. G. 1961 The development of proteolytie enzymes in the abomasum of the young calf. Proceedings of the Nutrition Society 20 xl–xliGoogle Scholar
Hsu, I. N., Delbaere, L. T. J., James, M. N. G. & Hofmann, T. 1977 Penicillopepsin from Penicillium janlhinellum crystal structure at 2·8 Å and sequence homology with porcine pepsin. Nature 266 140–145CrossRefGoogle ScholarPubMed
Jones, P. & Nicholson, B. H. 1979 Translational activity of RNA from ruminant abomasum. Biochemical Society Transactions 7 1253–1255CrossRefGoogle ScholarPubMed
Kessler, S. W. 1975 Rapid isolation of antigens from cells with a staphylococcal protein A–antibody adsorbent: parameters of the interaction of antibody–antigen complexes with protein A. Journal of Immunology 115 1617–1624CrossRefGoogle ScholarPubMed
Krystosek, A., Cawthon, M. L.. & Kabat, D. 1975 Improved methods for purification and assay of eukaryotic messenger ribonucleic acids and ribosomes: quantitative analysis of their interaction in a fractionated reticulocyte cell-free system. Journal of Biological Chemistry 250 6077–6084CrossRefGoogle Scholar
Laemmli, U. K. 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 680–685CrossRefGoogle ScholarPubMed
Lehraoh, H., Diamond, D., Wozney, J. M. & Boedtker, H. 1977 RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical re-examination. Biochemistry 16 4743–4751CrossRefGoogle Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. 1951 Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193 265–275CrossRefGoogle ScholarPubMed
Ouchterlony, Ö. 1964 Gel-diffusion techniques. In Immunological Methods, pp. 55–78 (Ed. Ackroyd, J. F.). Oxford: BlackwellGoogle Scholar
Palade, G. 1975 Intracellular aspects of the process of protein synthesis. Science 189 347–358CrossRefGoogle ScholarPubMed
Pelham, H. R. B. & Jackson, R. J. 1976 An efficient mRNA-dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67 247–256CrossRefGoogle ScholarPubMed
Roberts, B. E., Payne, P. I. & Osborne, D. J. 1973 Protein synthesis and the viability of rye grains. Loss of activity of protein-synthesizing systems in vitro associated with a loss of viability. Biochemical Journal 131 275–286CrossRefGoogle ScholarPubMed
Scheele, G., Dobberstein, B. & Blobel, G. 1978 Transfer of proteins across membranes. European Journl of Biochemistry 82 593–599CrossRefGoogle ScholarPubMed
Uchiyama, H., Uozumi, T., Beppu, T. & Arima, K. 1980 Purification of prorennin mRNA and its translation in vitro. Agricultural and Biological Chemistry 44 1373–1381Google Scholar
Yelton, D. E. & Scharff, M. D. 1981 Monoclonal antibodies: a powerful new tool in biology and medicine. Annual Review of Biochemistry 50 657–680CrossRefGoogle ScholarPubMed