Identification of the heparin-binding domain of TNF-alpha and its use for efficient TNF-alpha purification by heparin-Sepharose affinity chromatography

被引：20

作者：

Kenig, Maja ^{[1
]}

Gaberc-Porekar, Vladka ^{[2
]}

Fonda, Irena ^{[2
]}

Menart, Viktor ^{[1
,2
]}

机构：

[1] Lek Pharmaceut Dd, SL-1526 Ljubljana, Slovenia

[2] Natl Inst Chem, SL-1001 Ljubljana, Slovenia

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2008年 / 867卷 / 01期

关键词：

heparin-binding domain; TNF-alpha; affinity chromatography;

D O I：

10.1016/j.jchromb.2008.03.023

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The N-terminus of the trimeric TNF-alpha molecule comprises two basic arginines within the short amino-acid sequence VRSSSR, which is here shown to be essential for binding of TNF-alpha to heparin-Sepharose. Mixed trimers containing full-length and Delta N6-truncated subunits revealed a single VRSSSR sequence to be sufficient to achieve binding. On the basis of this newly identified heparin-binding domain, a new method for efficient purification of TNF-alpha is described. Affinity chromatography on heparin-Sepharose was introduced as a key step for highly purified TNF-alpha at a high yield. With minor modifications, this procedure can be used for TNF-alpha analogues that have full-length N-termini, as shown for the less toxic analogue LK-805. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：119 / 125

页数：7

共 36 条

[1]

ADAMS B, 1987, LYMPHOKINE RES, V6, P203

[2]

AGGARWAL BB, 1985, J BIOL CHEM, V260, P2345

[3] A novel human tumor necrosis factor alfa mutein, F4614, inhibits in vitro and in vivo growth of murine and human hepatoma:: Implication for immunotherapy of human hepatocellular carcinoma [J].

Atarashi, Y ;

Yasumura, S ;

Nambu, S ;

Yoshio, Y ;

Murakami, J ;

Takahara, T ;

Higuchi, K ;

Watanabe, A ;

Miyata, K ;

Kato, M .

HEPATOLOGY, 1998, 28 (01) :57-67

[4] DISSOCIATION OF TNF-ALPHA CYTOTOXIC AND PROINFLAMMATORY ACTIVITIES BY P55 RECEPTOR-SELECTIVE AND P75 RECEPTOR-SELECTIVE TNF-ALPHA MUTANTS [J].

BARBARA, JAJ ;

SMITH, WB ;

GAMBLE, JR ;

VANOSTADE, X ;

VANDENABEELE, P ;

TAVERNIER, J ;

FIERS, W ;

VADAS, MA ;

LOPEZ, AF .

EMBO JOURNAL, 1994, 13 (04) :843-850

[5]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[6] MOLECULAR MODELING OF PROTEIN-GLYCOSAMINOGLYCAN INTERACTIONS [J].

CARDIN, AD ;

WEINTRAUB, HJR .

ARTERIOSCLEROSIS, 1989, 9 (01) :21-32

[7]

*CFPMP EUR AG EV M, 1999, EUR PUBL ASS REP

[8] Identification of a major heparin-binding site in kallistatin [J].

Chen, VC ;

Chao, L ;

Pimenta, DC ;

Bledsoe, G ;

Juliano, L ;

Chao, J .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (02) :1276-1284

[9] STRUCTURE OF HUMAN-TUMOR NECROSIS FACTOR-ALPHA DERIVED FROM RECOMBINANT-DNA [J].

DAVIS, JM ;

NARACHI, MA ;

ALTON, NK ;

ARAKAWA, T .

BIOCHEMISTRY, 1987, 26 (05) :1322-1326

[10]

De Wilt JHW, 2000, ANTICANCER RES, V20, P3491

← 1 2 3 4 →