Comparative in vitro studies on native and recombinant human cationic trypsins: Cathepsin B is a possible pathological activator of trypsinogen in pancreatitis

László Szilágyi, Erzsébet Kénesi, Gergely Katona, Gyula Kaslik, G. Juhász, L. Gráf

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Hereditary pancreatitis, an autosomal dominant disease is believed to be caused by mutation in the human trypsinogen gene. The role of mutations has been investigated by in vitro studies using recombinant rat and human trypsinogen (TG). In this study we compare the enzymatic properties and inhibition by human pancreatic secretory trypsin inhibitor (hPSTI) of the native, postsynthetically modified and recombinant cationic trypsin, and found these values practically identical. We also determined the autolytic stability of recombinant wild type (Hu1Asn21) and pancreatitis-associated (Hu1Ile21) trypsin. Both forms were equally stable. Similarly, we found no difference in the rate of activation of the two zymogens by human cationic and anionic trypsin. Mesotrypsin did not activate either form. The rate of autocatalytic activation of Hu1Asn21 TG and Hu1Ile21 TG was also identical at pH 8 both in the presence and absence of Ca2+. At pH 5 Hu1Ile21 TG autoactivated about twice as fast as Hu1Asn21 TG. The presence of physiological amount of hPSTI completely prevented autoactivation of both zymogens at pH 8 and at pH 5 as well. Cathepsin B readily activated both zymogens although Hu1Ile21 TG was activated about 2.5-3 times as fast as Hu1Asn21 TG. The presence of hPSTI did not prevent the activation of zymogens by cathepsin B. Our results underlie the central role of cathepsin B in the development of different forms of pancreatitis.

Original languageEnglish
Pages (from-to)24574-24580
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number27
DOIs
Publication statusPublished - Jul 6 2001

Fingerprint

Trypsinogen
Cathepsin B
Pancreatitis
Trypsin
Enzyme Precursors
Kazal Pancreatic Trypsin Inhibitor
Chemical activation
Mutation
In Vitro Techniques
Rats
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Comparative in vitro studies on native and recombinant human cationic trypsins : Cathepsin B is a possible pathological activator of trypsinogen in pancreatitis. / Szilágyi, László; Kénesi, Erzsébet; Katona, Gergely; Kaslik, Gyula; Juhász, G.; Gráf, L.

In: Journal of Biological Chemistry, Vol. 276, No. 27, 06.07.2001, p. 24574-24580.

Research output: Contribution to journalArticle

@article{9b56ed8bda164a8b811963e758f9a3d8,
title = "Comparative in vitro studies on native and recombinant human cationic trypsins: Cathepsin B is a possible pathological activator of trypsinogen in pancreatitis",
abstract = "Hereditary pancreatitis, an autosomal dominant disease is believed to be caused by mutation in the human trypsinogen gene. The role of mutations has been investigated by in vitro studies using recombinant rat and human trypsinogen (TG). In this study we compare the enzymatic properties and inhibition by human pancreatic secretory trypsin inhibitor (hPSTI) of the native, postsynthetically modified and recombinant cationic trypsin, and found these values practically identical. We also determined the autolytic stability of recombinant wild type (Hu1Asn21) and pancreatitis-associated (Hu1Ile21) trypsin. Both forms were equally stable. Similarly, we found no difference in the rate of activation of the two zymogens by human cationic and anionic trypsin. Mesotrypsin did not activate either form. The rate of autocatalytic activation of Hu1Asn21 TG and Hu1Ile21 TG was also identical at pH 8 both in the presence and absence of Ca2+. At pH 5 Hu1Ile21 TG autoactivated about twice as fast as Hu1Asn21 TG. The presence of physiological amount of hPSTI completely prevented autoactivation of both zymogens at pH 8 and at pH 5 as well. Cathepsin B readily activated both zymogens although Hu1Ile21 TG was activated about 2.5-3 times as fast as Hu1Asn21 TG. The presence of hPSTI did not prevent the activation of zymogens by cathepsin B. Our results underlie the central role of cathepsin B in the development of different forms of pancreatitis.",
author = "L{\'a}szl{\'o} Szil{\'a}gyi and Erzs{\'e}bet K{\'e}nesi and Gergely Katona and Gyula Kaslik and G. Juh{\'a}sz and L. Gr{\'a}f",
year = "2001",
month = "7",
day = "6",
doi = "10.1074/jbc.M011374200",
language = "English",
volume = "276",
pages = "24574--24580",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "27",

}

TY - JOUR

T1 - Comparative in vitro studies on native and recombinant human cationic trypsins

T2 - Cathepsin B is a possible pathological activator of trypsinogen in pancreatitis

AU - Szilágyi, László

AU - Kénesi, Erzsébet

AU - Katona, Gergely

AU - Kaslik, Gyula

AU - Juhász, G.

AU - Gráf, L.

PY - 2001/7/6

Y1 - 2001/7/6

N2 - Hereditary pancreatitis, an autosomal dominant disease is believed to be caused by mutation in the human trypsinogen gene. The role of mutations has been investigated by in vitro studies using recombinant rat and human trypsinogen (TG). In this study we compare the enzymatic properties and inhibition by human pancreatic secretory trypsin inhibitor (hPSTI) of the native, postsynthetically modified and recombinant cationic trypsin, and found these values practically identical. We also determined the autolytic stability of recombinant wild type (Hu1Asn21) and pancreatitis-associated (Hu1Ile21) trypsin. Both forms were equally stable. Similarly, we found no difference in the rate of activation of the two zymogens by human cationic and anionic trypsin. Mesotrypsin did not activate either form. The rate of autocatalytic activation of Hu1Asn21 TG and Hu1Ile21 TG was also identical at pH 8 both in the presence and absence of Ca2+. At pH 5 Hu1Ile21 TG autoactivated about twice as fast as Hu1Asn21 TG. The presence of physiological amount of hPSTI completely prevented autoactivation of both zymogens at pH 8 and at pH 5 as well. Cathepsin B readily activated both zymogens although Hu1Ile21 TG was activated about 2.5-3 times as fast as Hu1Asn21 TG. The presence of hPSTI did not prevent the activation of zymogens by cathepsin B. Our results underlie the central role of cathepsin B in the development of different forms of pancreatitis.

AB - Hereditary pancreatitis, an autosomal dominant disease is believed to be caused by mutation in the human trypsinogen gene. The role of mutations has been investigated by in vitro studies using recombinant rat and human trypsinogen (TG). In this study we compare the enzymatic properties and inhibition by human pancreatic secretory trypsin inhibitor (hPSTI) of the native, postsynthetically modified and recombinant cationic trypsin, and found these values practically identical. We also determined the autolytic stability of recombinant wild type (Hu1Asn21) and pancreatitis-associated (Hu1Ile21) trypsin. Both forms were equally stable. Similarly, we found no difference in the rate of activation of the two zymogens by human cationic and anionic trypsin. Mesotrypsin did not activate either form. The rate of autocatalytic activation of Hu1Asn21 TG and Hu1Ile21 TG was also identical at pH 8 both in the presence and absence of Ca2+. At pH 5 Hu1Ile21 TG autoactivated about twice as fast as Hu1Asn21 TG. The presence of physiological amount of hPSTI completely prevented autoactivation of both zymogens at pH 8 and at pH 5 as well. Cathepsin B readily activated both zymogens although Hu1Ile21 TG was activated about 2.5-3 times as fast as Hu1Asn21 TG. The presence of hPSTI did not prevent the activation of zymogens by cathepsin B. Our results underlie the central role of cathepsin B in the development of different forms of pancreatitis.

UR - http://www.scopus.com/inward/record.url?scp=0035816544&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035816544&partnerID=8YFLogxK

U2 - 10.1074/jbc.M011374200

DO - 10.1074/jbc.M011374200

M3 - Article

C2 - 11312265

AN - SCOPUS:0035816544

VL - 276

SP - 24574

EP - 24580

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 27

ER -