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. 1991 Mar 1;112(5):981–989. doi: 10.1083/jcb.112.5.981

A photoreceptor calcium binding protein is recognized by autoantibodies obtained from patients with cancer-associated retinopathy

PMCID: PMC2288874  PMID: 1999465

Abstract

Cancer-associated retinopathy (CAR), a paraneoplastic syndrome, is characterized by the degeneration of retinal photoreceptors under conditions where the tumor and its metastases have not invaded the eye. The retinopathy often is apparent before the diagnosis of cancer and may be associated with autoantibodies that react with specific sites in the retina. We have examined the sera from patients with CAR to further characterize the retinal antigen. Western blot analysis of human retinal proteins reveals a prominent band at 26 kD that is labeled by the CAR antisera. Antibodies to the 26-kD protein were affinity- purified from complex CAR antisera and used for EM-immunocytochemical localization of the protein to the nuclei, inner and outer segments of both rod and cone cells. Other antibodies obtained from the CAR sera did not label photoreceptors. Using the affinity-purified antibodies for detection, the 26-kD protein, designated p26, was purified to homogeneity from the outer segments of bovine rod photoreceptor cells by Phenyl-Sepharose and ion exchange chromatography. Partial amino acid sequence of p26 was determined by gas phase Edman degradation and revealed extensive homology with a cone-specific protein, visinin. Based upon structural relatedness, both the p26 rod protein and visinin are members of the calmodulin family and contain calcium binding domains of the E-F hand structure.

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Selected References

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  1. Anderson N. E., Rosenblum M. K., Posner J. B. Paraneoplastic cerebellar degeneration: clinical-immunological correlations. Ann Neurol. 1988 Oct;24(4):559–567. doi: 10.1002/ana.410240413. [DOI] [PubMed] [Google Scholar]
  2. BRAIN W. R., DANIEL P. M., GREENFIELD J. G. Subacute cortical cerebellar degeneration and its relation to carcinoma. J Neurol Neurosurg Psychiatry. 1951 May;14(2):59–75. doi: 10.1136/jnnp.14.2.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Brain L., Wilkinson M. Subacute cerebellar degeneration associated with neoplasms. Brain. 1965 Sep;88(3):465–478. doi: 10.1093/brain/88.3.465. [DOI] [PubMed] [Google Scholar]
  5. Buchanan T. A., Gardiner T. A., Archer D. B. An ultrastructural study of retinal photoreceptor degeneration associated with bronchial carcinoma. Am J Ophthalmol. 1984 Mar;97(3):277–287. doi: 10.1016/0002-9394(84)90623-8. [DOI] [PubMed] [Google Scholar]
  6. Burton M. D., Onstott L. T., Polans A. S. The use of gold reagents to quantitate antibodies eluted from nitrocellulose blots: application to electron microscopic immunocytochemistry. Anal Biochem. 1989 Dec;183(2):225–230. doi: 10.1016/0003-2697(89)90472-7. [DOI] [PubMed] [Google Scholar]
  7. CRESTFIELD A. M., MOORE S., STEIN W. H. The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins. J Biol Chem. 1963 Feb;238:622–627. [PubMed] [Google Scholar]
  8. CROFT P. B., WILKINSON M. Carcinomatous neuromyopathy. Its incidence in patients with carcinoma of the lung and carcinoma of the breast. Lancet. 1963 Jan 26;1(7274):184–188. doi: 10.1016/s0140-6736(63)91208-x. [DOI] [PubMed] [Google Scholar]
  9. Coffee C. J., Bradshaw R. A. Carp muscle calcium-binding protein. I. Characterization of the tryptic peptides and the complete amino acid sequence of component B. J Biol Chem. 1973 May 10;248(9):3305–3312. [PubMed] [Google Scholar]
  10. Crabb J. W., Armes L. G., Carr S. A., Johnson C. M., Roberts G. D., Bordoli R. S., McKeehan W. L. Complete primary structure of prostatropin, a prostate epithelial cell growth factor. Biochemistry. 1986 Sep 9;25(18):4988–4993. doi: 10.1021/bi00366a003. [DOI] [PubMed] [Google Scholar]
  11. Crabb J. W., Johnson C. M., Carr S. A., Armes L. G., Saari J. C. The complete primary structure of the cellular retinaldehyde-binding protein from bovine retina. J Biol Chem. 1988 Dec 15;263(35):18678–18687. [PubMed] [Google Scholar]
  12. DENNY-BROWN D. Primary sensory neuropathy with muscular changes associated with carcinoma. J Neurol Neurosurg Psychiatry. 1948 May;11(2):73–87. doi: 10.1136/jnnp.11.2.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. De Aizpurua H. J., Lambert E. H., Griesmann G. E., Olivera B. M., Lennon V. A. Antagonism of voltage-gated calcium channels in small cell carcinomas of patients with and without Lambert-Eaton myasthenic syndrome by autoantibodies omega-conotoxin and adenosine. Cancer Res. 1988 Sep 1;48(17):4719–4724. [PubMed] [Google Scholar]
  14. Furneaux H. F., Reich L., Posner J. B. Autoantibody synthesis in the central nervous system of patients with paraneoplastic syndromes. Neurology. 1990 Jul;40(7):1085–1091. doi: 10.1212/wnl.40.7.1085. [DOI] [PubMed] [Google Scholar]
  15. Galbraith G. M., Emerson D., Fudenberg H. H., Gibbs C. J., Gajdusek D. C. Antibodies to neurofilament protein in retinitis pigmentosa. J Clin Invest. 1986 Oct;78(4):865–869. doi: 10.1172/JCI112672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Greenlee J. E., Lipton H. L. Anticerebellar antibodies in serum and cerebrospinal fluid of a patient with oat cell carcinoma of the lung and paraneoplastic cerebellar degeneration. Ann Neurol. 1986 Jan;19(1):82–85. doi: 10.1002/ana.410190117. [DOI] [PubMed] [Google Scholar]
  17. Grunwald G. B., Klein R., Simmonds M. A., Kornguth S. E. Autoimmune basis for visual paraneoplastic syndrome in patients with small-cell lung carcinoma. Lancet. 1985 Mar 23;1(8430):658–661. doi: 10.1016/s0140-6736(85)91326-1. [DOI] [PubMed] [Google Scholar]
  18. Hawley R. J., Cohen M. H., Saini N., Armbrustmacher V. W. The carcinomatous neuromyopathy of oat cell lung cancer. Ann Neurol. 1980 Jan;7(1):65–72. doi: 10.1002/ana.410070112. [DOI] [PubMed] [Google Scholar]
  19. Huang J. M., Matthews H. R. Application of sodium dodecyl sulfate-gel electrophoresis to low molecular weight polypeptides. Anal Biochem. 1990 Jul;188(1):114–117. doi: 10.1016/0003-2697(90)90537-j. [DOI] [PubMed] [Google Scholar]
  20. Johnson L. V., Blanks J. C. Application of acrylamide as an embedding medium in studies of lectin and antibody binding in the vertebrate retina. Curr Eye Res. 1984 Jul;3(7):969–974. doi: 10.3109/02713688409167215. [DOI] [PubMed] [Google Scholar]
  21. Keltner J. L., Roth A. M., Chang R. S. Photoreceptor degeneration. Possible autoimmune disorder. Arch Ophthalmol. 1983 Apr;101(4):564–569. doi: 10.1001/archopht.1983.01040010564006. [DOI] [PubMed] [Google Scholar]
  22. Kim Y. I., Neher E. IgG from patients with Lambert-Eaton syndrome blocks voltage-dependent calcium channels. Science. 1988 Jan 22;239(4838):405–408. doi: 10.1126/science.2447652. [DOI] [PubMed] [Google Scholar]
  23. Kornguth S. E., Klein R., Appen R., Choate J. Occurrence of anti-retinal ganglion cell antibodies in patients with small cell carcinoma of the lung. Cancer. 1982 Oct 1;50(7):1289–1293. doi: 10.1002/1097-0142(19821001)50:7<1289::aid-cncr2820500711>3.0.co;2-9. [DOI] [PubMed] [Google Scholar]
  24. Lai C. Y. Studies on the structure of rabbit muscle aldolase. I. Cleavage with cyanogen bromide: an approach to the determination of the total primary structure. Arch Biochem Biophys. 1968 Oct;128(1):201–211. [PubMed] [Google Scholar]
  25. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  26. Newsom-Davis J. Lambert-Eaton myasthenic syndrome. Springer Semin Immunopathol. 1985;8(1-2):129–140. doi: 10.1007/BF00197251. [DOI] [PubMed] [Google Scholar]
  27. Norris F. H., Jr The remote effects of cancer on the nervous system. Z Neurol. 1972;201(3):201–210. doi: 10.1007/BF00316412. [DOI] [PubMed] [Google Scholar]
  28. Papermaster D. S. Preparation of retinal rod outer segments. Methods Enzymol. 1982;81:48–52. doi: 10.1016/s0076-6879(82)81010-0. [DOI] [PubMed] [Google Scholar]
  29. Polans A. S., Burton M. D. Sialoglycoproteins of the frog rod outer segment plasma membrane. Invest Ophthalmol Vis Sci. 1988 Oct;29(10):1523–1532. [PubMed] [Google Scholar]
  30. Polans A. S., Hermolin J., Bownds M. D. Light-induced dephosphorylation of two proteins in frog rod outer segments: influence of cyclic nucleotides and calcium. J Gen Physiol. 1979 Nov;74(5):595–613. doi: 10.1085/jgp.74.5.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reinach F. C., Karlsson R. Cloning, expression, and site-directed mutagenesis of chicken skeletal muscle troponin C. J Biol Chem. 1988 Feb 15;263(5):2371–2376. [PubMed] [Google Scholar]
  32. Rogers J. H. Calretinin: a gene for a novel calcium-binding protein expressed principally in neurons. J Cell Biol. 1987 Sep;105(3):1343–1353. doi: 10.1083/jcb.105.3.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sawyer R. A., Selhorst J. B., Zimmerman L. E., Hoyt W. F. Blindness caused by photoreceptor degeneration as a remote effect of cancer. Am J Ophthalmol. 1976 May;81(5):606–613. doi: 10.1016/0002-9394(76)90125-2. [DOI] [PubMed] [Google Scholar]
  34. Schold S. C., Cho E. S., Somasundaram M., Posner J. B. Subacute motor neuronopathy: a remote effect of lymphoma. Ann Neurol. 1979 Mar;5(3):271–287. doi: 10.1002/ana.410050310. [DOI] [PubMed] [Google Scholar]
  35. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  36. Thirkill C. E., FitzGerald P., Sergott R. C., Roth A. M., Tyler N. K., Keltner J. L. Cancer-associated retinopathy (CAR syndrome) with antibodies reacting with retinal, optic-nerve, and cancer cells. N Engl J Med. 1989 Dec 7;321(23):1589–1594. doi: 10.1056/NEJM198912073212307. [DOI] [PubMed] [Google Scholar]
  37. Thirkill C. E., Roth A. M., Keltner J. L. Cancer-associated retinopathy. Arch Ophthalmol. 1987 Mar;105(3):372–375. doi: 10.1001/archopht.1987.01060030092033. [DOI] [PubMed] [Google Scholar]
  38. Tso M. O. Experiments on visual cells by nature and man: in search of treatment for photoreceptor degeneration. Friedenwald lecture. Invest Ophthalmol Vis Sci. 1989 Dec;30(12):2430–2454. [PubMed] [Google Scholar]
  39. Watterson D. M., Sharief F., Vanaman T. C. The complete amino acid sequence of the Ca2+-dependent modulator protein (calmodulin) of bovine brain. J Biol Chem. 1980 Feb 10;255(3):962–975. [PubMed] [Google Scholar]
  40. Wilden U., Kühn H. Light-dependent phosphorylation of rhodopsin: number of phosphorylation sites. Biochemistry. 1982 Jun 8;21(12):3014–3022. doi: 10.1021/bi00541a032. [DOI] [PubMed] [Google Scholar]
  41. Yamagata K., Goto K., Kuo C. H., Kondo H., Miki N. Visinin: a novel calcium binding protein expressed in retinal cone cells. Neuron. 1990 Mar;4(3):469–476. doi: 10.1016/0896-6273(90)90059-o. [DOI] [PubMed] [Google Scholar]

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