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

被引:79
作者
Szilágyi, L
Kénesi, E
Katona, G
Kaslik, G
Juhász, G
Gráf, L
机构
[1] Eotvos Lorand Univ, Dept Biochem, H-1088 Budapest, Hungary
[2] Hungarian Acad Sci, Biotechnol Res Grp, H-1088 Budapest, Hungary
[3] Eotvos Lorand Univ, Dept Physiol, H-1088 Budapest, Hungary
关键词
D O I
10.1074/jbc.M011374200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 HulAsn21 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.
引用
收藏
页码:24574 / 24580
页数:7
相关论文
共 35 条
[1]  
BRODRICK JW, 1978, J BIOL CHEM, V253, P2732
[2]   INVITRO MUTAGENESIS OF TRYPSINOGEN - ROLE OF THE AMINO TERMINUS IN INTRACELLULAR PROTEIN TARGETING TO SECRETORY GRANULES [J].
BURGESS, TL ;
CRAIK, CS ;
MATSUUCHI, L ;
KELLY, RB .
JOURNAL OF CELL BIOLOGY, 1987, 105 (02) :659-668
[3]  
Coleman P L, 1976, Methods Enzymol, V45, P12
[4]   2 HUMAN TRYPSINOGENS - CATALYTIC PROPERTIES OF CORRESPONDING TRYPSINS [J].
COLOMB, E ;
GUY, O ;
DEPREZ, P ;
MICHEL, R ;
FIGARELLA, C .
BIOCHIMICA ET BIOPHYSICA ACTA, 1978, 525 (01) :186-193
[5]   New PC versions of the kinetic-simulation and fitting programs, KINSIM and FITSIM [J].
Dang, Q ;
Frieden, C .
TRENDS IN BIOCHEMICAL SCIENCES, 1997, 22 (08) :317-317
[6]   THERMODYNAMICS AND KINETICS OF SINGLE RESIDUE REPLACEMENTS IN AVIAN OVOMUCOID 3RD DOMAINS - EFFECT ON INHIBITOR INTERACTIONS WITH SERINE PROTEINASES [J].
EMPIE, MW ;
LASKOWSKI, M .
BIOCHEMISTRY, 1982, 21 (10) :2274-2284
[7]   HUMAN PANCREATIC PROTEOLYTIC-ENZYMES AND PROTEIN INHIBITORS - ISOLATION AND MOLECULAR-PROPERTIES [J].
FEINSTEIN, G ;
HOFSTEIN, R ;
KOIFMANN, J ;
SOKOLOVSKY, M .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1974, 43 (03) :569-581
[8]  
FIGARELLA C, 1988, BIOL CHEM H-S, V369, P293
[9]   ADENOSINE-DEAMINASE AND ADENYLATE DEAMINASE - COMPARATIVE KINETIC-STUDIES WITH TRANSITION-STATE AND GROUND-STATE ANALOG INHIBITORS [J].
FRIEDEN, C ;
KURZ, LC ;
GILBERT, HR .
BIOCHEMISTRY, 1980, 19 (23) :5303-5309
[10]   Crystal structure of human trypsin 1: Unexpected phosphorylation of tyr151 [J].
Gaboriaud, C ;
Serre, L ;
GuyCrotte, O ;
Forest, E ;
FontecillaCamps, JC .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 259 (05) :995-1010