Stability towards alkaline conditions can be engineered into a protein ligand

被引：40

作者：

Gülich, S ^{[1
]}

Linhult, M ^{[1
]}

Nygren, PÅ ^{[1
]}

Uhlén, M ^{[1
]}

Hober, S ^{[1
]}

机构：

[1] Royal Inst Technol, Dept Biotechnol, SE-10044 Stockholm, Sweden

来源：

JOURNAL OF BIOTECHNOLOGY | 2000年 / 80卷 / 02期

关键词：

albumin-binding domain; cleaning-in-place; deamidation; human serum albumin; protein engineering;

D O I：

10.1016/S0168-1656(00)00259-5

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

One of the problems with a proteinaceous affinity ligand is their sensitivity to alkaline conditions. Here, we show that a simple and straightforward strategy consisting of replacing all asparagine residues with other amino acids can dramatically improve the chemical stability of a protein towards alkaline conditions. As a model, a Streptococcal albumin-binding domain (ABD) was used. The engineered variant showed higher stability towards 0.5 M NaOH, as well as higher thermal stability compared to its native counterpart. This protein engineering approach could potentially also be used for other protein ligands to eliminate the sensitivity to alkaline cleaning-in-place (CIP) conditions. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：169 / 178

页数：10

共 17 条

[1]

DECHATEAU M, 1994, J BIOL CHEM, V269, P12147

[2]

GEIGER T, 1987, J BIOL CHEM, V262, P785

[3] A GENERAL-METHOD OF INVITRO PREPARATION AND SPECIFIC MUTAGENESIS OF DNA FRAGMENTS - STUDY OF PROTEIN AND DNA INTERACTIONS [J].

HIGUCHI, R ;

KRUMMEL, B ;

SAIKI, RK .

NUCLEIC ACIDS RESEARCH, 1988, 16 (15) :7351-7367

[4]

JOHNSON CW, 1990, PROTEIN-STRUCT FUNCT, V7, P205

[5] TERTIARY STRUCTURE IS A PRINCIPAL DETERMINANT TO PROTEIN DEAMIDATION [J].

KOSSIAKOFF, AA .

SCIENCE, 1988, 240 (4849) :191-194

[6] The serum albumin-binding domain of streptococcal protein G is a three-helical bundle: A heteronuclear NMR study [J].

Kraulis, PJ ;

Jonasson, P ;

Nygren, PA ;

Uhlen, M ;

Jendeberg, L ;

Nilsson, B ;

Kordel, J .

FEBS LETTERS, 1996, 378 (02) :190-194

[7] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[8] Design, synthesis, and application of a Protein A mimetic [J].

Li, RX ;

Dowd, V ;

Stewart, DJ ;

Burton, SJ ;

Lowe, CR .

NATURE BIOTECHNOLOGY, 1998, 16 (02) :190-195

[9] ROLE OF PEPTIDE CONFORMATION IN THE RATE AND MECHANISM OF DEAMIDATION OF ASPARAGINYL RESIDUES [J].

LURA, R ;

SCHIRCH, V .

BIOCHEMISTRY, 1988, 27 (20) :7671-7677

[10] STRUCTURE AND EVOLUTION OF THE REPETITIVE GENE ENCODING STREPTOCOCCAL PROTEIN-G [J].

OLSSON, A ;

ELIASSON, M ;

GUSS, B ;

NILSSON, B ;

HELLMAN, U ;

LINDBERG, M ;

UHLEN, M .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1987, 168 (02) :319-324

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