Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Apr 1;117(1):121–133. doi: 10.1083/jcb.117.1.121

Tissue and cellular distribution of the extended family of protein kinase C isoenzymes

PMCID: PMC2289401  PMID: 1556149

Abstract

Polyclonal isoenzyme-specific antisera were developed against four calcium-independent protein kinase C (PKC) isoenzymes (delta, epsilon, epsilon', and zeta) as well as the calcium-dependent isoforms (alpha, beta I, beta II, and gamma). These antisera showed high specificities, high titers, and high binding affinities (3-370 nM) for the peptide antigens to which they were raised. Each antiserum detected a species of the predicted molecular weight by Western blot that could be blocked with the immunizing peptide. PKC was sequentially purified from rat brain, and the calcium-dependent forms were finally resolved by hydroxyapatite chromatography. Peak I reacted exclusively with antisera to PKC gamma, peak II with PKC beta I and -beta II, and peak III with PKC alpha. These same fractions, however, were devoid of immunoreactivity for the calcium-independent isoenzymes. The PKC isoenzymes demonstrated a distinctive tissue distribution when evaluated by Western blot and immunocytochemistry. PCK delta was present in brain, heart, spleen, lung, liver, ovary, pancreas, and adrenal tissues. PKC epsilon was present in brain, kidney, and pancreas, whereas PKC epsilon' was present predominantly in brain. PKC zeta was present in most tissues, particularly the lung, brain, and liver. Both PKC delta and PKC zeta showed some heterogeneity of size among the different tissues. PKC alpha was present in all organs and tissues examined. PKC beta I and -beta II were present in greatest amount in brain and spleen. Although the brain contained the most PKC gamma immunoreactivity, some immunostaining was also seen in adrenal tissue. These studies provide the first evidence of selective organ and tissue distributions of the calcium-independent PKC isoenzymes.

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

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

  1. Akita Y., Ohno S., Konno Y., Yano A., Suzuki K. Expression and properties of two distinct classes of the phorbol ester receptor family, four conventional protein kinase C types, and a novel protein kinase C. J Biol Chem. 1990 Jan 5;265(1):354–362. [PubMed] [Google Scholar]
  2. Ase K., Saito N., Shearman M. S., Kikkawa U., Ono Y., Igarashi K., Tanaka C., Nishizuka Y. Distinct cellular expression of beta I- and beta II-subspecies of protein kinase C in rat cerebellum. J Neurosci. 1988 Oct;8(10):3850–3856. doi: 10.1523/JNEUROSCI.08-10-03850.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bacher N., Zisman Y., Berent E., Livneh E. Isolation and characterization of PKC-L, a new member of the protein kinase C-related gene family specifically expressed in lung, skin, and heart. Mol Cell Biol. 1991 Jan;11(1):126–133. doi: 10.1128/mcb.11.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bell R. M. Protein kinase C activation by diacylglycerol second messengers. Cell. 1986 Jun 6;45(5):631–632. doi: 10.1016/0092-8674(86)90774-9. [DOI] [PubMed] [Google Scholar]
  5. 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Brandt S. J., Niedel J. E., Bell R. M., Young W. S., 3rd Distinct patterns of expression of different protein kinase C mRNAs in rat tissues. Cell. 1987 Apr 10;49(1):57–63. doi: 10.1016/0092-8674(87)90755-0. [DOI] [PubMed] [Google Scholar]
  7. Burns D. J., Bloomenthal J., Lee M. H., Bell R. M. Expression of the alpha, beta II, and gamma protein kinase C isozymes in the baculovirus-insect cell expression system. Purification and characterization of the individual isoforms. J Biol Chem. 1990 Jul 15;265(20):12044–12051. [PubMed] [Google Scholar]
  8. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  9. Coussens L., Parker P. J., Rhee L., Yang-Feng T. L., Chen E., Waterfield M. D., Francke U., Ullrich A. Multiple, distinct forms of bovine and human protein kinase C suggest diversity in cellular signaling pathways. Science. 1986 Aug 22;233(4766):859–866. doi: 10.1126/science.3755548. [DOI] [PubMed] [Google Scholar]
  10. Culler M. D., Negro-Vilar A. Development of specific antisera and a radioimmunoassay procedure for the gonadotropin-releasing hormone associated peptide (GAP) of the LHRH prohormone. Brain Res Bull. 1986 Aug;17(2):219–223. doi: 10.1016/0361-9230(86)90118-8. [DOI] [PubMed] [Google Scholar]
  11. Hannun Y. A., Loomis C. R., Bell R. M. Activation of protein kinase C by Triton X-100 mixed micelles containing diacylglycerol and phosphatidylserine. J Biol Chem. 1985 Aug 25;260(18):10039–10043. [PubMed] [Google Scholar]
  12. Hashimoto K., Kishimoto A., Aihara H., Yasuda I., Mikawa K., Nishizuka Y. Protein kinase C during differentiation of human promyelocytic leukemia cell line, HL-60. FEBS Lett. 1990 Apr 9;263(1):31–34. doi: 10.1016/0014-5793(90)80698-i. [DOI] [PubMed] [Google Scholar]
  13. Hidaka H., Tanaka T., Onoda K., Hagiwara M., Watanabe M., Ohta H., Ito Y., Tsurudome M., Yoshida T. Cell type-specific expression of protein kinase C isozymes in the rabbit cerebellum. J Biol Chem. 1988 Apr 5;263(10):4523–4526. [PubMed] [Google Scholar]
  14. Huang F. L., Yoshida Y., Nakabayashi H., Huang K. P. Differential distribution of protein kinase C isozymes in the various regions of brain. J Biol Chem. 1987 Nov 15;262(32):15714–15720. [PubMed] [Google Scholar]
  15. Huang F. L., Yoshida Y., Nakabayashi H., Knopf J. L., Young W. S., 3rd, Huang K. P. Immunochemical identification of protein kinase C isozymes as products of discrete genes. Biochem Biophys Res Commun. 1987 Dec 31;149(3):946–952. doi: 10.1016/0006-291x(87)90500-6. [DOI] [PubMed] [Google Scholar]
  16. Huang F. L., Yoshida Y., Nakabayashi H., Young W. S., 3rd, Huang K. P. Immunocytochemical localization of protein kinase C isozymes in rat brain. J Neurosci. 1988 Dec;8(12):4734–4744. doi: 10.1523/JNEUROSCI.08-12-04734.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huang K. P., Huang F. L., Nakabayashi H., Yoshida Y. Biochemical characterization of rat brain protein kinase C isozymes. J Biol Chem. 1988 Oct 15;263(29):14839–14845. [PubMed] [Google Scholar]
  18. Huang K. P., Nakabayashi H., Huang F. L. Isozymic forms of rat brain Ca2+-activated and phospholipid-dependent protein kinase. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8535–8539. doi: 10.1073/pnas.83.22.8535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ito A., Saito N., Taniguchi H., Chiba T., Kikkawa U., Saitoh Y., Tanaka C. Localization of beta II subspecies of protein kinase C in beta-cells. Diabetes. 1989 Aug;38(8):1005–1011. doi: 10.2337/diab.38.8.1005. [DOI] [PubMed] [Google Scholar]
  20. Kikkawa U., Ogita K., Ono Y., Asaoka Y., Shearman M. S., Fujii T., Ase K., Sekiguchi K., Igarashi K., Nishizuka Y. The common structure and activities of four subspecies of rat brain protein kinase C family. FEBS Lett. 1987 Nov 2;223(2):212–216. doi: 10.1016/0014-5793(87)80291-0. [DOI] [PubMed] [Google Scholar]
  21. Kiley S., Schaap D., Parker P., Hsieh L. L., Jaken S. Protein kinase C heterogeneity in GH4C1 rat pituitary cells. Characterization of a Ca2(+)-independent phorbol ester receptor. J Biol Chem. 1990 Sep 15;265(26):15704–15712. [PubMed] [Google Scholar]
  22. Kitano T., Go M., Kikkawa U., Nishizuka Y. Assay and purification of protein kinase C. Methods Enzymol. 1986;124:349–352. doi: 10.1016/0076-6879(86)24026-4. [DOI] [PubMed] [Google Scholar]
  23. Knopf J. L., Lee M. H., Sultzman L. A., Kriz R. W., Loomis C. R., Hewick R. M., Bell R. M. Cloning and expression of multiple protein kinase C cDNAs. Cell. 1986 Aug 15;46(4):491–502. doi: 10.1016/0092-8674(86)90874-3. [DOI] [PubMed] [Google Scholar]
  24. Konno Y., Ohno S., Akita Y., Kawasaki H., Suzuki K. Enzymatic properties of a novel phorbol ester receptor/protein kinase, nPKC. J Biochem. 1989 Oct;106(4):673–678. doi: 10.1093/oxfordjournals.jbchem.a122915. [DOI] [PubMed] [Google Scholar]
  25. Kosaka Y., Ogita K., Ase K., Nomura H., Kikkawa U., Nishizuka Y. The heterogeneity of protein kinase C in various rat tissues. Biochem Biophys Res Commun. 1988 Mar 30;151(3):973–981. doi: 10.1016/s0006-291x(88)80461-3. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Leach K. L., Powers E. A., McGuire J. C., Dong L., Kiley S. C., Jaken S. Monoclonal antibodies specific for type 3 protein kinase C recognize distinct domains of protein kinase C and inhibit in vitro functional activity. J Biol Chem. 1988 Sep 15;263(26):13223–13230. [PubMed] [Google Scholar]
  28. Lowenthal J. W., Ballard D. W., Bogerd H., Böhnlein E., Greene W. C. Tumor necrosis factor-alpha activation of the IL-2 receptor-alpha gene involves the induction of kappa B-specific DNA binding proteins. J Immunol. 1989 May 1;142(9):3121–3128. [PubMed] [Google Scholar]
  29. Makowske M., Ballester R., Cayre Y., Rosen O. M. Immunochemical evidence that three protein kinase C isozymes increase in abundance during HL-60 differentiation induced by dimethyl sulfoxide and retinoic acid. J Biol Chem. 1988 Mar 5;263(7):3402–3410. [PubMed] [Google Scholar]
  30. Mavis R. D., Bell R. M., Vagelos P. R. Effect of phospholipase C hydrolysis of membrane phospholipids on membranous enzymes. J Biol Chem. 1972 May 10;247(9):2835–2841. [PubMed] [Google Scholar]
  31. Merchenthaler I., Vigh S., Petrusz P., Schally A. V. Immunocytochemical localization of corticotropin-releasing factor (CRF) in the rat brain. Am J Anat. 1982 Dec;165(4):385–396. doi: 10.1002/aja.1001650404. [DOI] [PubMed] [Google Scholar]
  32. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  34. Nishizuka Y. Studies and prospectives of the protein kinase c family for cellular regulation. Cancer. 1989 May 15;63(10):1892–1903. doi: 10.1002/1097-0142(19890515)63:10<1892::aid-cncr2820631005>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  35. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  36. Ohno S., Akita Y., Konno Y., Imajoh S., Suzuki K. A novel phorbol ester receptor/protein kinase, nPKC, distantly related to the protein kinase C family. Cell. 1988 Jun 3;53(5):731–741. doi: 10.1016/0092-8674(88)90091-8. [DOI] [PubMed] [Google Scholar]
  37. Ohno S., Kawasaki H., Imajoh S., Suzuki K., Inagaki M., Yokokura H., Sakoh T., Hidaka H. Tissue-specific expression of three distinct types of rabbit protein kinase C. Nature. 1987 Jan 8;325(7000):161–166. doi: 10.1038/325161a0. [DOI] [PubMed] [Google Scholar]
  38. Ono Y., Fujii T., Igarashi K., Kuno T., Tanaka C., Kikkawa U., Nishizuka Y. Phorbol ester binding to protein kinase C requires a cysteine-rich zinc-finger-like sequence. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4868–4871. doi: 10.1073/pnas.86.13.4868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ono Y., Fujii T., Ogita K., Kikkawa U., Igarashi K., Nishizuka Y. Protein kinase C zeta subspecies from rat brain: its structure, expression, and properties. Proc Natl Acad Sci U S A. 1989 May;86(9):3099–3103. doi: 10.1073/pnas.86.9.3099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ono Y., Fujii T., Ogita K., Kikkawa U., Igarashi K., Nishizuka Y. The structure, expression, and properties of additional members of the protein kinase C family. J Biol Chem. 1988 May 15;263(14):6927–6932. [PubMed] [Google Scholar]
  41. Onoda K., Hagiwara M., Hachiya T., Usuda N., Nagata T., Hidaka H. Different expression of protein kinase C isozymes in pancreatic islet cells. Endocrinology. 1990 Feb;126(2):1235–1240. doi: 10.1210/endo-126-2-1235. [DOI] [PubMed] [Google Scholar]
  42. Ordronneau P., Lindström P. B., Petrusz P. Four unlabeled antibody bridge techniques: a comparison. J Histochem Cytochem. 1981 Dec;29(12):1397–1404. doi: 10.1177/29.12.7033366. [DOI] [PubMed] [Google Scholar]
  43. Osada S., Mizuno K., Saido T. C., Akita Y., Suzuki K., Kuroki T., Ohno S. A phorbol ester receptor/protein kinase, nPKC eta, a new member of the protein kinase C family predominantly expressed in lung and skin. J Biol Chem. 1990 Dec 25;265(36):22434–22440. [PubMed] [Google Scholar]
  44. Pelosin J. M., Ricouart A., Sergheraert C., Benahmed M., Chambaz E. M. Expression of protein kinase C isoforms in various steroidogenic cell types. Mol Cell Endocrinol. 1991 Feb;75(2):149–155. doi: 10.1016/0303-7207(91)90229-l. [DOI] [PubMed] [Google Scholar]
  45. Saito N., Kose A., Ito A., Hosoda K., Mori M., Hirata M., Ogita K., Kikkawa U., Ono Y., Igarashi K. Immunocytochemical localization of beta II subspecies of protein kinase C in rat brain. Proc Natl Acad Sci U S A. 1989 May;86(9):3409–3413. doi: 10.1073/pnas.86.9.3409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sawamura S., Ase K., Berry N., Kikkawa U., McCaffrey P. G., Minowada J., Nishizuka Y. Expression of protein kinase C subspecies in human leukemia-lymphoma cell lines. FEBS Lett. 1989 Apr 24;247(2):353–357. doi: 10.1016/0014-5793(89)81369-9. [DOI] [PubMed] [Google Scholar]
  47. Schaap D., Hsuan J., Totty N., Parker P. J. Proteolytic activation of protein kinase C-epsilon. Eur J Biochem. 1990 Jul 31;191(2):431–435. doi: 10.1111/j.1432-1033.1990.tb19139.x. [DOI] [PubMed] [Google Scholar]
  48. Schaap D., Parker P. J., Bristol A., Kriz R., Knopf J. Unique substrate specificity and regulatory properties of PKC-epsilon: a rationale for diversity. FEBS Lett. 1989 Jan 30;243(2):351–357. doi: 10.1016/0014-5793(89)80160-7. [DOI] [PubMed] [Google Scholar]
  49. Shearman M. S., Naor Z., Kikkawa U., Nishizuka Y. Differential expression of multiple protein kinase C subspecies in rat central nervous tissue. Biochem Biophys Res Commun. 1987 Sep 30;147(3):911–919. doi: 10.1016/s0006-291x(87)80157-2. [DOI] [PubMed] [Google Scholar]
  50. Shimohama S., Saitoh T., Gage F. H. Differential expression of protein kinase C isozymes in rat cerebellum. J Chem Neuroanat. 1990 Sep-Oct;3(5):367–375. [PubMed] [Google Scholar]
  51. 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]
  52. Tsujino T., Kose A., Saito N., Tanaka C. Light and electron microscopic localization of beta I-, beta II-, and gamma-subspecies of protein kinase C in rat cerebral neocortex. J Neurosci. 1990 Mar;10(3):870–884. doi: 10.1523/JNEUROSCI.10-03-00870.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Watanabe M., Hachiya T., Hagiwara M., Hidaka H. Identification of type III protein kinase C in bovine aortic tissue. Arch Biochem Biophys. 1989 Aug 15;273(1):165–169. doi: 10.1016/0003-9861(89)90175-6. [DOI] [PubMed] [Google Scholar]
  54. Yoshida Y., Huang F. L., Nakabayashi H., Huang K. P. Tissue distribution and developmental expression of protein kinase C isozymes. J Biol Chem. 1988 Jul 15;263(20):9868–9873. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES