Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1995 Apr 15;307(Pt 2):481–486. doi: 10.1042/bj3070481

Molecular cloning, purification and in situ localization of human colon kallikrein.

L M Chen 1, G P Richards 1, L Chao 1, J Chao 1
PMCID: PMC1136673  PMID: 7733886

Abstract

We have cloned and characterized a full-length cDNA encoding tissue kallikrein from a human colon carcinoma cell line (T84). The nucleic acid sequence of the colon kallikrein cDNA is identical to that of renal/pancreatic or tissue kallikrein cDNA. Reverse-transcription PCR followed by Southern-blot analysis using specific oligonucleotide probes showed expression of tissue kallikrein in human colon, pancreas and kidney. Tissue kallikrein mRNA was localized in glandular epithelial cells (goblet cells) in colon by in situ hybridization histochemistry. Human colon kallikrein was purified to apparent homogeneity by DEAE-Sepharose Cl-6B, aprotinin-affinity, and HQ/M perfusion chromatography. The purified colon kallikrein migrated as a broad, 40-45 kDa band in SDS/PAGE and was recognized by antibodies to human tissue kallikrein. The linear displacement curves for the colon kallikrein in an RIA were parallel with the human tissue kallikrein standard curve, indicating their immunological identity. The N-terminal sequence of the purified colon kallikrein matches completely with that of purified urinary or tissue kallikrein. These results indicate that human colon kallikrein is transcribed from the tissue kallikrein gene.

Full text

PDF
481

Images in this article

483Figure 1
on p.483
484Figure 2
on p.484
484Figure 3
on p.484

Selected References

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

  1. Ashley P. L., MacDonald R. J. Kallikrein-related mRNAs of the rat submaxillary gland: nucleotide sequences of four distinct types including tonin. Biochemistry. 1985 Aug 13;24(17):4512–4520. doi: 10.1021/bi00338a005. [DOI] [PubMed] [Google Scholar]
  2. Baird A. W., Miller D. H., Schwartz D. A., Margolius H. S. Enhancement of kallikrein production and kinin sensitivity in T84 cells by growth in the nude mouse. Am J Physiol. 1991 Nov;261(5 Pt 1):C822–C827. doi: 10.1152/ajpcell.1991.261.5.C822. [DOI] [PubMed] [Google Scholar]
  3. Chapdelaine P., Paradis G., Tremblay R. R., Dubé J. Y. High level of expression in the prostate of a human glandular kallikrein mRNA related to prostate-specific antigen. FEBS Lett. 1988 Aug 15;236(1):205–208. doi: 10.1016/0014-5793(88)80315-6. [DOI] [PubMed] [Google Scholar]
  4. Clements J. A. The glandular kallikrein family of enzymes: tissue-specific expression and hormonal regulation. Endocr Rev. 1989 Nov;10(4):393–419. doi: 10.1210/edrv-10-4-393. [DOI] [PubMed] [Google Scholar]
  5. Cuthbert A. W., MacVinish L. J. Transporting epithelia as targets for kinin effects. Adv Exp Med Biol. 1986;198(Pt A):203–210. doi: 10.1007/978-1-4684-5143-6_28. [DOI] [PubMed] [Google Scholar]
  6. Cuthbert A. W., Margolius H. S. Kinins stimulate net chloride secretion by the rat colon. Br J Pharmacol. 1982 Apr;75(4):587–598. doi: 10.1111/j.1476-5381.1982.tb09178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans B. A., Yun Z. X., Close J. A., Tregear G. W., Kitamura N., Nakanishi S., Callen D. F., Baker E., Hyland V. J., Sutherland G. R. Structure and chromosomal localization of the human renal kallikrein gene. Biochemistry. 1988 May 3;27(9):3124–3129. doi: 10.1021/bi00409a003. [DOI] [PubMed] [Google Scholar]
  8. Fukushima D., Kitamura N., Nakanishi S. Nucleotide sequence of cloned cDNA for human pancreatic kallikrein. Biochemistry. 1985 Dec 31;24(27):8037–8043. doi: 10.1021/bi00348a030. [DOI] [PubMed] [Google Scholar]
  9. Fuller P. J., Verity K., Matheson B. A., Clements J. A. Kallikrein-gene expression in the rat gastrointestinal tract. Biochem J. 1989 Nov 15;264(1):133–136. doi: 10.1042/bj2640133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerald W. L., Chao J., Chao L. Immunological identification of rat tissue kallikrein cDNA and characterization of the kallikrein gene family. Biochim Biophys Acta. 1986 Feb 24;866(1):1–14. doi: 10.1016/0167-4781(86)90093-x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Lin F. K., Lin C. H., Chou C. C., Chen K., Lu H. S., Bacheller W., Herrera C., Jones T., Chao J., Chao L. Molecular cloning and sequence analysis of the monkey and human tissue kallikrein genes. Biochim Biophys Acta. 1993 Jun 25;1173(3):325–328. doi: 10.1016/0167-4781(93)90131-v. [DOI] [PubMed] [Google Scholar]
  13. Lu H. S., Lin F. K., Chao L., Chao J. Human urinary kallikrein. Complete amino acid sequence and sites of glycosylation. Int J Pept Protein Res. 1989 Apr;33(4):237–249. doi: 10.1111/j.1399-3011.1989.tb01277.x. [DOI] [PubMed] [Google Scholar]
  14. MacDonald R. J., Margolius H. S., Erdös E. G. Molecular biology of tissue kallikrein. Biochem J. 1988 Jul 15;253(2):313–321. doi: 10.1042/bj2530313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mason A. J., Evans B. A., Cox D. R., Shine J., Richards R. I. Structure of mouse kallikrein gene family suggests a role in specific processing of biologically active peptides. Nature. 1983 May 26;303(5915):300–307. doi: 10.1038/303300a0. [DOI] [PubMed] [Google Scholar]
  16. Morita T., Kato H., Iwanaga S., Takada K., Kimura T. New fluorogenic substrates for alpha-thrombin, factor Xa, kallikreins, and urokinase. J Biochem. 1977 Nov;82(5):1495–1498. doi: 10.1093/oxfordjournals.jbchem.a131840. [DOI] [PubMed] [Google Scholar]
  17. Morris B. J. hGK-1: a kallikrein gene expressed in human prostate. Clin Exp Pharmacol Physiol. 1989 Apr;16(4):345–351. doi: 10.1111/j.1440-1681.1989.tb01570.x. [DOI] [PubMed] [Google Scholar]
  18. Murray S. R., Chao J., Lin F. K., Chao L. Kallikrein multigene families and the regulation of their expression. J Cardiovasc Pharmacol. 1990;15 (Suppl 6):S7–16. [PubMed] [Google Scholar]
  19. Riegman P. H., Vlietstra R. J., Suurmeijer L., Cleutjens C. B., Trapman J. Characterization of the human kallikrein locus. Genomics. 1992 Sep;14(1):6–11. doi: 10.1016/s0888-7543(05)80275-7. [DOI] [PubMed] [Google Scholar]
  20. Schachter M. Kallikreins (kininogenases)--a group of serine proteases with bioregulatory actions. Pharmacol Rev. 1979 Mar;31(1):1–17. [PubMed] [Google Scholar]
  21. Schachter M., Longridge D. J., Wheeler G. D., Mehta J. G., Uchida Y. Immunocytochemical and enzyme histochemical localization of kallikrein-like enzymes in colon, intestine, and stomach of rat and cat. J Histochem Cytochem. 1986 Jul;34(7):927–934. doi: 10.1177/34.7.3519756. [DOI] [PubMed] [Google Scholar]
  22. Schachter M., Peret M. W., Billing A. G., Wheeler G. D. Immunolocalization of the protease kallikrein in the colon. J Histochem Cytochem. 1983 Nov;31(11):1255–1260. doi: 10.1177/31.11.6194202. [DOI] [PubMed] [Google Scholar]
  23. Schedlich L. J., Bennetts B. H., Morris B. J. Primary structure of a human glandular kallikrein gene. DNA. 1987 Oct;6(5):429–437. doi: 10.1089/dna.1987.6.429. [DOI] [PubMed] [Google Scholar]
  24. Shimamoto K., Chao J., Margolius H. S. The radioimmunoassay of human urinary kallikrein and comparisons with kallikrein activity measurements. J Clin Endocrinol Metab. 1980 Oct;51(4):840–848. doi: 10.1210/jcem-51-4-840. [DOI] [PubMed] [Google Scholar]
  25. Skagen D. W., Andersen K. J. Intestinal kallikrein activity is reduced in a bypassed segment of the small intestine in the rat. Comp Biochem Physiol A Comp Physiol. 1986;84(1):175–180. doi: 10.1016/0300-9629(86)90060-5. [DOI] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. Sutherland G. R., Baker E., Hyland V. J., Callen D. F., Close J. A., Tregear G. W., Evans B. A., Richards R. I. Human prostate-specific antigen (APS) is a member of the glandular kallikrein gene family at 19q13. Cytogenet Cell Genet. 1988;48(4):205–207. doi: 10.1159/000132629. [DOI] [PubMed] [Google Scholar]
  28. Wines D. R., Brady J. M., Pritchett D. B., Roberts J. L., MacDonald R. J. Organization and expression of the rat kallikrein gene family. J Biol Chem. 1989 May 5;264(13):7653–7662. [PubMed] [Google Scholar]
  29. Zimmermann A., Geiger R., Kortmann H. Similarity between a kininogenase (kallikrein) from human large intestine and human urinary kallikrein. Hoppe Seylers Z Physiol Chem. 1979 Dec;360(12):1767–1773. doi: 10.1515/bchm2.1979.360.2.1767. [DOI] [PubMed] [Google Scholar]

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

RESOURCES