Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1981 May 15;196(2):653–656. doi: 10.1042/bj1960653

Identification of Limulus polyphemus haemocyanin messenger RNA.

E J Wood, J Bonaventura
PMCID: PMC1163044  PMID: 7317005

Abstract

Total RNA was isolated from cyanoblast-containing tissue taken from behind the compound eye of the horseshoe crab, Limulus polyphemus. Poly(A)-containing RNA separated from this by affinity chromatography on oligo(dT)-cellulose was translated in the rabbit reticulocyte haemolysate system in the presence of L-[35S]methionine. By using an antiserum to Limulus haemocyanin, polypeptides were isolated from the translation products which had a similar mobility to the authentic Limulus haemocyanin polypeptides as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis.

Full text

PDF
653

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amara S. G., Rosenfeld M. G., Birnbaum R. S., Roos B. A. Identification of the putative cell-free translation product of rat calcitonin mRNA. J Biol Chem. 1980 Apr 10;255(7):2645–2648. [PubMed] [Google Scholar]
  2. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blobel G., Dobberstein B. Transfer of proteins across membranes. II. Reconstitution of functional rough microsomes from heterologous components. J Cell Biol. 1975 Dec;67(3):852–862. doi: 10.1083/jcb.67.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Craig R. K., Boulton A. P., Harrison O. S., Parker D., Campbell P. N. Studies on the intracellular segregation of polyribosome-associated messenger ribonucleic acid species in the lactating guinea-pig mammary gland. Biochem J. 1979 Sep 1;181(3):737–756. doi: 10.1042/bj1810737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dilly P. N., Messenger J. B. The branchial gland: a site of haemocyanin synthesis in Octopus. Z Zellforsch Mikrosk Anat. 1972;132(2):193–201. doi: 10.1007/BF00307010. [DOI] [PubMed] [Google Scholar]
  7. Evans G. A., Rosenfeld M. G. Cell-free synthesis of a prolactin precursor directed by mRNA from cultured rat pituitary cells. J Biol Chem. 1976 May 10;251(9):2842–2847. [PubMed] [Google Scholar]
  8. Fahrenbach W. H. The cyanoblast: hemocyanin formation in Limulus polyphemus. J Cell Biol. 1970 Feb;44(2):445–453. doi: 10.1083/jcb.44.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ghiretti-Magaldi A., Milanesi C., Salvato B. Identification of hemocyanin in the cyanocytes of Carcinus maenas. Experientia. 1973 Oct 15;29(10):1265–1267. doi: 10.1007/BF01935109. [DOI] [PubMed] [Google Scholar]
  10. Gielens C., Verschueren L. J., Préaux G., Lontie R. Fragmentation of crystalline beta-haemocyanin of Helix pomatia with plasmin and trypsin. Location of the fragments in the polypeptide chain. Eur J Biochem. 1980 Feb;103(3):463–470. doi: 10.1111/j.1432-1033.1980.tb05970.x. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lamy J., Lamy J., Weill J., Bonaventura J., Bonaventura C., Brenowitz M. Immunological correlates between the multiple hemocyanin subunits of Limulus polyphemus and Techypleus tridentatus. Arch Biochem Biophys. 1979 Sep;196(2):324–339. doi: 10.1016/0003-9861(79)90585-x. [DOI] [PubMed] [Google Scholar]
  13. Payvar F., Schimke R. T. Methylmercury hydroxide enhancement of translation and transcription of ovalbumin and conalbumin mRNA's. J Biol Chem. 1979 Aug 25;254(16):7636–7642. [PubMed] [Google Scholar]
  14. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  15. Sullivan B., Bonaventura J., Bonaventura C., Godette G. Hemocyanin of the horseshoe crab, Limulus polyphemus. Structural differentiation of the isolated components. J Biol Chem. 1976 Dec 10;251(23):7644–7648. [PubMed] [Google Scholar]
  16. Wood E. J., Siggens K. W., Hall R. L., Orton J. M. Isolation of poly(adenosine) containing ribonucleic acid from the tissues of gastropod molluscs and its translation in a heterologous system [proceedings]. Biochem Soc Trans. 1979 Apr;7(2):389–390. doi: 10.1042/bst0070389. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES