Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Mar 15;346(Pt 3):817–826.

Latexin, a carboxypeptidase A inhibitor, is expressed in rat peritoneal mast cells and is associated with granular structures distinct from secretory granules and lysosomes.

Y Uratani 1, K Takiguchi-Hayashi 1, N Miyasaka 1, M Sato 1, M Jin 1, Y Arimatsu 1
PMCID: PMC1220918  PMID: 10698712

Abstract

Latexin, a protein possessing inhibitory activity against rat carboxypeptidase A1 (CPA1) and CPA2, is expressed in a neuronal subset in the cerebral cortex and cells in other neural and non-neural tissues of rat. Although latexin also inhibits mast-cell CPA (MCCPA), the expression of latexin in rat mast cells has not previously been confirmed. In the present study we examined the expression and subcellular localization of latexin in rat peritoneal mast cells. Western blot and reverse-transcriptase-mediated PCR analyses showed that latexin was contained and expressed in the rat peritoneal mast cells. Immunocytochemically, latexin immunofluorescence was localized on granular structures distinct from MCCPA-, histamine- or cathepsin D-immunopositive granules. Immunoelectron microscopy revealed that latexin was associated with a minority population of granules. The latexin-associated granules were separated from MCCPA- or histamine-containing granules on a self-generating density gradient of polyvinylpyrrolidone-coated silica-gel particles (Percoll). Treatments with high ionic strength and heparinase released latexin from the granules, suggesting that latexin is non-covalently associated with a heparin-like component of the granules. MCCPA and histamine were released from the mast cells after non-immunological and immunological stimulation with compound 48/80, A23187 and anti-IgE antibody, whereas latexin was not released. These results show that latexin is synthesized in rat peritoneal mast cells and suggest that it is associated with a unique type of intracellular granules distinct from MCCPA- and histamine-containing secretory granules and lysosomes.

Full Text

The Full Text of this article is available as a PDF (288.0 KB).

Selected References

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

  1. Akiyama M., Watanabe Y., Nishikawa T. Immunohistochemical characterization of human cutaneous mast cells in urticaria pigmentosa (cutaneous mastocytosis) Acta Pathol Jpn. 1991 May;41(5):344–349. doi: 10.1111/j.1440-1827.1991.tb01657.x. [DOI] [PubMed] [Google Scholar]
  2. Arimatsu Y., Miyamoto M., Nihonmatsu I., Hirata K., Uratani Y., Hatanaka Y., Takiguchi-Hayashi K. Early regional specification for a molecular neuronal phenotype in the rat neocortex. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8879–8883. doi: 10.1073/pnas.89.19.8879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arimatsu Y., Nihonmatsu I., Hirata K., Takiguchi-Hayashi K. Cogeneration of neurons with a unique molecular phenotype in layers V and VI of widespread lateral neocortical areas in the rat. J Neurosci. 1994 Apr;14(4):2020–2031. doi: 10.1523/JNEUROSCI.14-04-02020.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baram D., Adachi R., Medalia O., Tuvim M., Dickey B. F., Mekori Y. A., Sagi-Eisenberg R. Synaptotagmin II negatively regulates Ca2+-triggered exocytosis of lysosomes in mast cells. J Exp Med. 1999 May 17;189(10):1649–1658. doi: 10.1084/jem.189.10.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clauser E., Gardell S. J., Craik C. S., MacDonald R. J., Rutter W. J. Structural characterization of the rat carboxypeptidase A1 and B genes. Comparative analysis of the rat carboxypeptidase gene family. J Biol Chem. 1988 Nov 25;263(33):17837–17845. [PubMed] [Google Scholar]
  6. Cole K. R., Kumar S., Trong H. L., Woodbury R. G., Walsh K. A., Neurath H. Rat mast cell carboxypeptidase: amino acid sequence and evidence of enzyme activity within mast cell granules. Biochemistry. 1991 Jan 22;30(3):648–655. doi: 10.1021/bi00217a009. [DOI] [PubMed] [Google Scholar]
  7. Everitt M. T., Neurath H. Rat peritoneal mast cell carboxypeptidase: localization, purification, and enzymatic properties. FEBS Lett. 1980 Feb 11;110(2):292–296. doi: 10.1016/0014-5793(80)80095-0. [DOI] [PubMed] [Google Scholar]
  8. Fiorucci L., Erba F., Falasca L., Dini L., Ascoli F. Localization and interaction of bovine pancreatic trypsin inhibitor and tryptase in the granules of bovine mast cells. Biochim Biophys Acta. 1995 Apr 13;1243(3):407–413. doi: 10.1016/0304-4165(94)00167-v. [DOI] [PubMed] [Google Scholar]
  9. Gardell S. J., Craik C. S., Clauser E., Goldsmith E. J., Stewart C. B., Graf M., Rutter W. J. A novel rat carboxypeptidase, CPA2: characterization, molecular cloning, and evolutionary implications on substrate specificity in the carboxypeptidase gene family. J Biol Chem. 1988 Nov 25;263(33):17828–17836. [PubMed] [Google Scholar]
  10. Harvima I. T., Haapanen L., Ackermann L., Naukkarinen A., Harvima R. J., Horsmanheimo M. Decreased chymase activity is associated with increased levels of protease inhibitors in mast cells of psoriatic lesions. Acta Derm Venereol. 1999 Mar;79(2):98–104. doi: 10.1080/000155599750011282. [DOI] [PubMed] [Google Scholar]
  11. Harvima I. T., Naukkarinen A., Paukkonen K., Harvima R. J., Aalto M. L., Schwartz L. B., Horsmanheimo M. Mast cell tryptase and chymase in developing and mature psoriatic lesions. Arch Dermatol Res. 1993;285(4):184–192. doi: 10.1007/BF00372007. [DOI] [PubMed] [Google Scholar]
  12. Hatanaka Y., Uratani Y., Takiguchi-Hayashi K., Omori A., Sato K., Miyamoto M., Arimatsu Y. Intracortical regionality represented by specific transcription for a novel protein, latexin. Eur J Neurosci. 1994 Jun 1;6(6):973–982. doi: 10.1111/j.1460-9568.1994.tb00592.x. [DOI] [PubMed] [Google Scholar]
  13. Itoh H., Ide H., Ishikawa N., Nawa Y. Mast cell protease inhibitor, trypstatin, is a fragment of inter-alpha-trypsin inhibitor light chain. J Biol Chem. 1994 Feb 4;269(5):3818–3822. [PubMed] [Google Scholar]
  14. Jamur M. C., Vugman I., Hand A. R. Ultrastructural and cytochemical studies of acid phosphatase and trimetaphosphatase in rat peritoneal mast cells developing in vivo. Cell Tissue Res. 1986;244(3):557–563. doi: 10.1007/BF00212533. [DOI] [PubMed] [Google Scholar]
  15. Kaminska R., Harvima I. T., Naukkarinen A., Nilsson G., Horsmanheimo M. Alterations in mast cell proteinases and protease inhibitors in the progress of cutaneous herpes zoster infection. J Pathol. 1996 Dec;180(4):434–440. doi: 10.1002/(SICI)1096-9896(199612)180:4<434::AID-PATH682>3.0.CO;2-B. [DOI] [PubMed] [Google Scholar]
  16. Kido H., Yokogoshi Y., Katunuma N. Kunitz-type protease inhibitor found in rat mast cells. Purification, properties, and amino acid sequence. J Biol Chem. 1988 Dec 5;263(34):18104–18107. [PubMed] [Google Scholar]
  17. Koffer A., Tatham P. E., Gomperts B. D. Changes in the state of actin during the exocytotic reaction of permeabilized rat mast cells. J Cell Biol. 1990 Sep;111(3):919–927. doi: 10.1083/jcb.111.3.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koibuchi Y., Ichikawa A., Nakagawa M., Tomita K. Histamine release induced from mast cells by active components of compound 48/80. Eur J Pharmacol. 1985 Sep 24;115(2-3):163–170. doi: 10.1016/0014-2999(85)90687-9. [DOI] [PubMed] [Google Scholar]
  19. Krüger P. G., Lagunoff D., Wan H. Isolation of rat mast cell granules with intact membranes. Exp Cell Res. 1980 Sep;129(1):83–93. doi: 10.1016/0014-4827(80)90333-x. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. 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]
  22. Lawson D., Raff M. C., Gomperts B., Fewtrell C., Gilula N. B. Molecular events during membrane fusion. A study of exocytosis in rat peritoneal mast cells. J Cell Biol. 1977 Feb;72(2):242–259. doi: 10.1083/jcb.72.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lindahl U., Lidholt K., Spillmann D., Kjellén L. More to "heparin" than anticoagulation. Thromb Res. 1994 Jul 1;75(1):1–32. doi: 10.1016/0049-3848(94)90136-8. [DOI] [PubMed] [Google Scholar]
  24. Lützelschwab C., Pejler G., Aveskogh M., Hellman L. Secretory granule proteases in rat mast cells. Cloning of 10 different serine proteases and a carboxypeptidase A from various rat mast cell populations. J Exp Med. 1997 Jan 6;185(1):13–29. doi: 10.1084/jem.185.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miyasaka N., Hatanaka Y., Jin M., Arimatsu Y. Genomic organization and regulatory elements of the rat latexin gene, which is expressed in a cell type-specific manner in both central and peripheral nervous systems. Brain Res Mol Brain Res. 1999 May 21;69(1):62–72. doi: 10.1016/s0169-328x(99)00107-2. [DOI] [PubMed] [Google Scholar]
  26. Morrison D. C., Roser J. F., Henson P. M., Cochrane C. G. Activation of rat mast cells by low molecular weight stimuli. J Immunol. 1974 Feb;112(2):573–582. [PubMed] [Google Scholar]
  27. Muno D., Ishidoh K., Ueno T., Kominami E. Processing and transport of the precursor of cathepsin C during its transfer into lysosomes. Arch Biochem Biophys. 1993 Oct;306(1):103–110. doi: 10.1006/abbi.1993.1486. [DOI] [PubMed] [Google Scholar]
  28. Normant E., Gros C., Schwartz J. C. Carboxypeptidase A isoforms produced by distinct genes or alternative splicing in brain and other extrapancreatic tissues. J Biol Chem. 1995 Sep 1;270(35):20543–20549. doi: 10.1074/jbc.270.35.20543. [DOI] [PubMed] [Google Scholar]
  29. Normant E., Martres M. P., Schwartz J. C., Gros C. Purification, cDNA cloning, functional expression, and characterization of a 26-kDa endogenous mammalian carboxypeptidase inhibitor. Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12225–12229. doi: 10.1073/pnas.92.26.12225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pallaoro M., Gambacurta A., Fiorucci L., Mignogna G., Barra D., Ascoli F. cDNA cloning and primary structure of tryptase from bovine mast cells, and evidence for the expression of bovine pancreatic trypsin inhibitor mRNA in the same cells. Eur J Biochem. 1996 Apr 1;237(1):100–105. doi: 10.1111/j.1432-1033.1996.0100t.x. [DOI] [PubMed] [Google Scholar]
  31. SHORE P. A., BURKHALTER A., COHN V. H., Jr A method for the fluorometric assay of histamine in tissues. J Pharmacol Exp Ther. 1959 Nov;127:182–186. [PubMed] [Google Scholar]
  32. Sannes P. L., McDonald J. K., Allen R. C., Spicer S. S. Cytochemical localization and biochemical characterization of dipeptidyl aminopeptidase II in macrophages and mast cells. J Histochem Cytochem. 1979 Nov;27(11):1496–1498. doi: 10.1177/27.11.512331. [DOI] [PubMed] [Google Scholar]
  33. Schueller E., Peutsch M., Bohacek L. G., Gupta R. K. A simplified toluidine blue stain for mast cells. Can J Med Technol. 1967 Aug;29(4):137–138. [PubMed] [Google Scholar]
  34. Schwartz L. B., Riedel C., Schratz J. J., Austen K. F. Localization of carboxypeptidase A to the macromolecular heparin proteoglycan-protein complex in secretory granules of rat serosal mast cells. J Immunol. 1982 Mar;128(3):1128–1133. [PubMed] [Google Scholar]
  35. Takiguchi-Hayashi K., Arimatsu Y. Restricted expression of latexin in dorsal midline cells of developing rat forebrain. Neuroreport. 1995 Jan 26;6(2):281–283. doi: 10.1097/00001756-199501000-00014. [DOI] [PubMed] [Google Scholar]
  36. Takiguchi-Hayashi K., Sato M., Sugo N., Ishida M., Sato K., Uratani Y., Arimatsu Y. Latexin expression in smaller diameter primary sensory neurons in the rat. Brain Res. 1998 Aug 10;801(1-2):9–20. doi: 10.1016/s0006-8993(98)00496-x. [DOI] [PubMed] [Google Scholar]
  37. Uratani Y. Immunoaffinity purification and reconstitution of sodium-coupled branched-chain amino acid carrier of Pseudomonas aeruginosa. J Biol Chem. 1992 Mar 15;267(8):5177–5183. [PubMed] [Google Scholar]
  38. Westin U., Polling A., Ljungkrantz I., Ohlsson K. Identification of SLPI (secretory leukocyte protease inhibitor) in human mast cells using immunohistochemistry and in situ hybridisation. Biol Chem. 1999 Apr;380(4):489–493. doi: 10.1515/BC.1999.063. [DOI] [PubMed] [Google Scholar]

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

RESOURCES