An immobilized biotin ligase:: Surface display of Escherichia coli BirA on Saccharomyces cerevisiae

被引：13

作者：

Parthasarathy, R

Bajaj, J

Boder, ET

机构：

[1] Univ Penn, Dept Chem & Biomol Engn, Philadelphia, PA 19104 USA

[2] Univ Penn, Res Struct Matter Lab, Philadelphia, PA 19104 USA

来源：

BIOTECHNOLOGY PROGRESS | 2005年 / 21卷 / 06期

关键词：

D O I：

10.1021/bp050279t

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：

The Escherichia coli biotin ligase enzyme BirA has been extensively used in recent years to generate site-specifically biotinylated proteins via a biotin acceptor peptide tag. In the present study, BirA was displayed for the first time on the yeast Saccharomyces cerevisiae using the Aga1p-Aga2p platform and assayed using a peptide-tagged protein as the substrate. The enzyme is fully functional and resembles the soluble form in many of its properties, but the yeast-displayed enzyme demonstrates stability and reusability on the time scale of weeks. Thus, the yeast-displayed BirA system represents a facile and highly economical alternative for producing site-specifically biotinylated proteins.

引用

收藏

页码：1627 / 1631

页数：5

相关论文

共 18 条

[1] Phenotypic analysis of antigen-specific T lymphocytes [J].

Altman, JD ;

Moss, PAH ;

Goulder, PJR ;

Barouch, DH ;

McHeyzerWilliams, MG ;

Bell, JI ;

McMichael, AJ ;

Davis, MM .

SCIENCE, 1996, 274 (5284) :94-96

[2] Control of gene expression with small molecules: Biotin-mediated acylation of targeted lysine residues in recombinant yeast [J].

Athavankar, S ;

Peterson, BR .

CHEMISTRY & BIOLOGY, 2003, 10 (12) :1245-1253

[3] A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation [J].

Beckett, D ;

Kovaleva, E ;

Schatz, PJ .

PROTEIN SCIENCE, 1999, 8 (04) :921-929

[4] Yeast surface display for directed evolution of protein expression, affinity, and stability [J].

Boder, ET ;

Wittrup, KD .

APPLICATIONS OF CHIMERIC GENES AND HYBRID PROTEINS, PT C: PROTEIN-PROTEIN INTERACTIONS AND GENOMICS, 2000, 328 :430-444

[5]

CAPELLARO C, 1991, EMBO J, V10, P4081

[6] The enzymatic biotinylation of proteins: a post-translational modification of exceptional specificity [J].

Chapman-Smith, A ;

Cronan, JE .

TRENDS IN BIOCHEMICAL SCIENCES, 1999, 24 (09) :359-363

[7] Detection of antigen-specific T cells with multivalent soluble class II MHC covalent peptide complexes [J].

Crawford, F ;

Kozono, H ;

White, J ;

Marrack, P ;

Kappler, J .

IMMUNITY, 1998, 8 (06) :675-682

[8]

Cull MG, 2000, METHOD ENZYMOL, V326, P430

[9] Efficient biotinylation and single-step purification of tagged transcription factors in mammalian cells and transgenic mice [J].

de Boer, E ;

Rodriguez, P ;

Bonte, E ;

Krijgsveld, J ;

Katsantoni, E ;

Heck, A ;

Grosveld, F ;

Strouboulis, J .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (13) :7480-7485

[10]

Gietz RD, 2002, METHOD ENZYMOL, V350, P87