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. 1992 Oct 15;89(20):9584–9587. doi: 10.1073/pnas.89.20.9584

Imaging of luciferase secretion from transformed Chinese hamster ovary cells.

S Inouye 1, Y Ohmiya 1, Y Toya 1, F I Tsuji 1
PMCID: PMC50176  PMID: 1409669

Abstract

The blue luminescence characteristic of the marine ostracod crustacean Vargula hilgendorfii is from a simple, but highly specific, enzyme-substrate reaction. Light is emitted by the oxidation of Vargula luciferin (substrate) by molecular oxygen, a reaction catalyzed by luciferase. Stable transformants of Chinese hamster ovary cells carrying the Vargula luciferase gene secreted luciferase from discrete sites on the cell surface, and this secretion could be monitored in real time by the bioluminescence produced by the secreted luciferase in the presence of Vargula luciferin by using an image-intensifying technique. Addition of anti-Vargula luciferase IgG to the luminescing cells almost completely extinguished the luminescence, confirming that Vargula luciferase caused the luminescence.

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Selected References

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

  1. Burgess T. L., Kelly R. B. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. doi: 10.1146/annurev.cb.03.110187.001331. [DOI] [PubMed] [Google Scholar]
  2. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FEDDEN G. A., CHASE A. M. The diffusion constant of Cypridina luciferase. Biochim Biophys Acta. 1959 Mar;32(1):176–181. doi: 10.1016/0006-3002(59)90566-9. [DOI] [PubMed] [Google Scholar]
  4. Hunziker W., Whitney J. A., Mellman I. Selective inhibition of transcytosis by brefeldin A in MDCK cells. Cell. 1991 Nov 1;67(3):617–627. doi: 10.1016/0092-8674(91)90535-7. [DOI] [PubMed] [Google Scholar]
  5. Inouye S., Guo Y., DePaolo L., Shimonaka M., Ling N., Shimasaki S. Recombinant expression of human follistatin with 315 and 288 amino acids: chemical and biological comparison with native porcine follistatin. Endocrinology. 1991 Aug;129(2):815–822. doi: 10.1210/endo-129-2-815. [DOI] [PubMed] [Google Scholar]
  6. Inouye S., Tsuji F. I. Monitoring gene expression in Chinese hamster ovary cells using secreted apoaequorin. Anal Biochem. 1992 Feb 14;201(1):114–118. doi: 10.1016/0003-2697(92)90182-7. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Lippincott-Schwartz J., Yuan L., Tipper C., Amherdt M., Orci L., Klausner R. D. Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic. Cell. 1991 Nov 1;67(3):601–616. doi: 10.1016/0092-8674(91)90534-6. [DOI] [PubMed] [Google Scholar]
  9. Lynch R. V., 3rd, Tsuji F. I., Donald D. H. Evidence for a calcium requirement in the Cypridina bioluminescence reaction. Biochem Biophys Res Commun. 1972 Feb 25;46(4):1544–1550. doi: 10.1016/0006-291x(72)90783-8. [DOI] [PubMed] [Google Scholar]
  10. Orci L., Tagaya M., Amherdt M., Perrelet A., Donaldson J. G., Lippincott-Schwartz J., Klausner R. D., Rothman J. E. Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin-coated buds on Golgi cisternae. Cell. 1991 Mar 22;64(6):1183–1195. doi: 10.1016/0092-8674(91)90273-2. [DOI] [PubMed] [Google Scholar]
  11. Pelham H. R. Multiple targets for brefeldin A. Cell. 1991 Nov 1;67(3):449–451. doi: 10.1016/0092-8674(91)90517-3. [DOI] [PubMed] [Google Scholar]
  12. SHIMOMURA O., JOHNSON F. H., SAIGA Y. Purification and properties of Cypridina luciferase. J Cell Comp Physiol. 1961 Oct;58:113–123. doi: 10.1002/jcp.1030580202. [DOI] [PubMed] [Google Scholar]
  13. Shimomura O., Johnson F. H., Masugi T. Cypridina bioluminescence: light-emitting oxyluciferin-luciferase complex. Science. 1969 Jun 13;164(3885):1299–1300. doi: 10.1126/science.164.3885.1299. [DOI] [PubMed] [Google Scholar]
  14. Shimomura O., Johnson F. H. Mechanism of the luminescent oxidation of cypridina luciferin. Biochem Biophys Res Commun. 1971 Jul 16;44(2):340–346. doi: 10.1016/0006-291x(71)90605-x. [DOI] [PubMed] [Google Scholar]
  15. Stone H. The enzyme catalyzed oxidation of Cypridina luciferin. Biochem Biophys Res Commun. 1968 May 10;31(3):386–391. doi: 10.1016/0006-291x(68)90487-7. [DOI] [PubMed] [Google Scholar]
  16. Subramani S., Mulligan R., Berg P. Expression of the mouse dihydrofolate reductase complementary deoxyribonucleic acid in simian virus 40 vectors. Mol Cell Biol. 1981 Sep;1(9):854–864. doi: 10.1128/mcb.1.9.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sugiura S., Inoue S., Goto T. [Synthesis of Cypridina luciferin and related compounds. 8. Improvement in the synthesis of Cypridina luciferin]. Yakugaku Zasshi. 1970 Jun;90(6):707–710. doi: 10.1248/yakushi1947.90.6_707. [DOI] [PubMed] [Google Scholar]
  18. TSUJI F. I., DAVIS D. L. A quantitative photometric method for studying the reaction between Cypridina luciferase and specific antibody. J Immunol. 1959 Feb;82(2):153–160. [PubMed] [Google Scholar]
  19. TSUJI F. I., DAVIS D. L., GINDLER E. M. Effect of sodium chloride and pH on the rate of neutralization of Cypridina luciferase by specific antibody. J Immunol. 1962 Jan;88:83–92. [PubMed] [Google Scholar]
  20. TSUJI F. I., DAVIS D. L., SOWINSKI R. Kinetics of the inhibition of Cypridina luciferase by specific antibody. J Immunol. 1960 Jun;84:615–625. [PubMed] [Google Scholar]
  21. Thompson E. M., Nagata S., Tsuji F. I. Cloning and expression of cDNA for the luciferase from the marine ostracod Vargula hilgendorfii. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6567–6571. doi: 10.1073/pnas.86.17.6567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thompson E. M., Nagata S., Tsuji F. I. Vargula hilgendorfii luciferase: a secreted reporter enzyme for monitoring gene expression in mammalian cells. Gene. 1990 Dec 15;96(2):257–262. [PubMed] [Google Scholar]
  23. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tsuji F. I., Davis D. L., Donald D. H. The effect of hydrostatic pressure on the rate of inactivation of Cypridina luciferase by specific antibody. J Immunol. 1966 Apr;96(4):614–621. [PubMed] [Google Scholar]
  25. Tsuji F. I., Lynch R. V., 3rd, Stevens C. L. Some properties of luciferase from the bioluminescent crustacean, Cypridina hilgendorfii. Biochemistry. 1974 Dec 3;13(25):5204–5209. doi: 10.1021/bi00722a024. [DOI] [PubMed] [Google Scholar]
  26. Urlaub G., Chasin L. A. Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4216–4220. doi: 10.1073/pnas.77.7.4216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wood S. A., Park J. E., Brown W. J. Brefeldin A causes a microtubule-mediated fusion of the trans-Golgi network and early endosomes. Cell. 1991 Nov 1;67(3):591–600. doi: 10.1016/0092-8674(91)90533-5. [DOI] [PubMed] [Google Scholar]

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