A rapid and sensitive method to measure the enzymatic activity of ribosome-inactivating proteins

被引：41

作者：

Brigotti, M ^{[1
]}

Barbieri, L ^{[1
]}

Valbonesi, P ^{[1
]}

Stirpe, F ^{[1
]}

Montanaro, L ^{[1
]}

Sperti, S ^{[1
]}

机构：

[1] Univ Bologna, Dipartimento Patol Sperimentale, I-40126 Bologna, Italy

来源：

NUCLEIC ACIDS RESEARCH | 1998年 / 26卷 / 18期

关键词：

D O I：

10.1093/nar/26.18.4306

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：

A method is described in which the adenosine-N-glycosidase activity of ribosome-inactivating proteins (RIPs) is measured using as substrate a 2251 bp [H-3]DNA obtained by PCR amplification of the 731-2981 region of the pBR322 plasmid, The DNA, labelled in the purine ring of adenine, proved a good substrate for all three RIPs tested (PAP-S, ricin and shiga-like toxin I). The method, which measures directly the [H-3]adenine released, is highly specific, extremely rapid and quantitative in a wide range of RIP concentrations.

引用

页码：4306 / 4307

页数：2

共 8 条

[1] Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins: Effect on DNA, RNA and poly(A) [J].

Barbieri, L ;

Valbonesi, P ;

Bonora, E ;

Gorini, P ;

Bolognesi, A ;

Stirpe, F .

NUCLEIC ACIDS RESEARCH, 1997, 25 (03) :518-522

[2] RIBOSOME-INACTIVATING PROTEINS FROM PLANTS [J].

BARBIERI, L ;

BATTELLI, MG ;

STIRPE, F .

BIOCHIMICA ET BIOPHYSICA ACTA, 1993, 1154 (3-4) :237-282

[3]

BARBIERI L, 1998, IN PRESS MOL MICROBI

[4] The RNA-N-glycosidase activity of shiga-like toxin I: Kinetic parameters of the native and activated toxin [J].

Brigotti, M ;

Carnicelli, D ;

Alvergna, P ;

Mazzaracchio, R ;

Sperti, S ;

Montanaro, L .

TOXICON, 1997, 35 (09) :1431-1437

[5]

ENDO Y, 1988, J BIOL CHEM, V263, P8735

[6]

ENDO Y, 1987, J BIOL CHEM, V262, P5908

[7] RIBOSOME INACTIVATION BY TOXIC LECTINS ABRIN AND RICIN - KINETICS OF ENZYMIC ACTIVITY OF TOXIN A-CHAINS [J].

OLSNES, S ;

FERNANDEZPUENTES, C ;

CARRASCO, L ;

VAZQUEZ, D .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1975, 60 (01) :281-288

[8] HIGH-PRESSURE-LIQUID-CHROMATOGRAPHIC AND FLUORIMETRIC METHODS FOR THE DETERMINATION OF ADENINE RELEASED FROM RIBOSOMES BY RICIN AND GELONIN [J].

ZAMBONI, M ;

BRIGOTTI, M ;

RAMBELLI, F ;

MONTANARO, L ;

SPERTI, S .

BIOCHEMICAL JOURNAL, 1989, 259 (03) :639-643

← 1 →