High-level expression and purification of a nonmitogenic form of human acidic fibroblast growth factor in Escherichia coli

被引：36

作者：

Wu, XP ^{[1
]}

Su, ZJ ^{[1
]}

Li, XK ^{[1
]}

Zheng, Q ^{[1
]}

Huang, YD ^{[1
]}

Yuan, H ^{[1
]}

机构：

[1] Jinan Univ, Coll Pharm, Biopharmaceut Res & Dev Ctr, Guangzhou 510632, Peoples R China

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2005年 / 42卷 / 01期

关键词：

nonmitogenic; human acidic fibroblast growth factor; purification;

D O I：

10.1016/j.pep.2004.07.021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

To decrease the potential side effects of acidic fibroblast growth factor (aFGF) caused by its broad-spectrum mitogenic activity, a nonmitogenic form of aFGF (nhaFGF), which retained the cardio- and neuroprotective characters of the wild-type aFGF, was over-expressed in Escherichia coli. The expression level of nhaFGF was up to 25% of the total cellular protein. The expressed nhaFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. The mitogenic activity of the purified nhaFGF was decreased dramatically comparable to that of the wild-type aFGF (haFGF) detected by methylthiazoletetrazolium method. The purified recombinant nhaFGF was sufficiently prepared and sufficient for the following pharmacological study. (c) 2004 Elsevier Inc. All rights reserved.

引用

页码：7 / 11

页数：5

共 9 条

[1] THE IMPORTANCE OF ARG40 AND ARG45 IN THE MITOGENIC ACTIVITY AND STRUCTURAL STABILITY OF BASIC FIBROBLAST GROWTH-FACTOR - EFFECTS OF ACIDIC AMINO-ACID SUBSTITUTIONS [J].

ARAKAWA, T ;

HOLST, P ;

NARHI, LO ;

PHILO, JS ;

WEN, J ;

PRESTRELSKI, SJ ;

ZHU, XT ;

REES, DC ;

FOX, GM .

JOURNAL OF PROTEIN CHEMISTRY, 1995, 14 (05) :263-274

[2] Protection of rat myocardium by mitogenic and non-mitogenic fibroblast growth factor during post-ischemic reperfusion [J].

Cuevas, P ;

Carceller, F ;

Lozano, RM ;

Crespo, A ;

Zazo, M ;

GimenezGallego, G .

GROWTH FACTORS, 1997, 15 (01) :29-+

[3] Intravenous fibroblast growth factor penetrates the blood-brain barrier and protects hippocampal neurons against ischemia-reperfusion injury [J].

Cuevas, P ;

Carceller, F ;

Munoz-Willery, I ;

Gimenez-Gallego, G .

SURGICAL NEUROLOGY, 1998, 49 (01) :77-83

[4]

GIMENEZGALLEGO G, 1994, NEUROL RES, V16, P313

[5]

LI X, 2001, J S CHINA U TECHNOLO, V29, P61

[6] 1H NMR structural characterization of a nonmitogenic, vasodilatory, ischemia-protector and neuromodulatory acidic fibroblast growth factor [J].

Lozano, RM ;

Pineda-Lucena, A ;

Gonzalez, C ;

Jiménez, MA ;

Cuevas, P ;

Redondo-Horcajo, M ;

Sanz, JM ;

Rico, M ;

Giménez-Gallego, G .

BIOCHEMISTRY, 2000, 39 (17) :4982-4993

[7] Three-dimensional structure of acidic fibroblast growth factor in solution: Effects of binding to a heparin functional analog [J].

PinedaLucena, A ;

Jimenez, MA ;

Lozano, RM ;

Nieto, JL ;

Santoro, J ;

Rico, M ;

GimenezGallego, G .

JOURNAL OF MOLECULAR BIOLOGY, 1996, 264 (01) :162-178

[8] X-ray structure of native full-length human fibroblast-growth factor at 0.25-nm resolution [J].

Romero, A ;

PinedaLucena, A ;

GimenezGallego, G .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1996, 241 (02) :453-461

[9]

SPRINGER BA, 1994, J BIOL CHEM, V269, P26879

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