BirA enzyme: Production and application in the study of membrane receptor-ligand interactions by site-specific biotinylation

被引:94
作者
O'Callaghan, CA [1 ]
Byford, MF
Wyer, JR
Willcox, BE
Jakobsen, BK
McMichael, AJ
Bell, JI
机构
[1] Univ Oxford, John Radcliffe Hosp, Nuffield Dept Clin Med, Mol Immunol Grp,Inst Mol Med, Oxford OX3 9DS, England
[2] Univ Oxford, Oxford Ctr Mol Sci, Dyson Perrins Lab, Oxford OX1 3QY, England
关键词
BirA; biotin; streptavidin; membrane proteins;
D O I
10.1006/abio.1998.2930
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The enzyme BirA is a key reagent because of its ability to biotinylate proteins at a specific residue in a recognition sequence. We report a rapid, efficient, and economical method for the production, purification, and application of this enzyme, The method is easily scaled up and the protein produced is of high purity and can be stored for many months with retention of activity. We have used this enzyme to biotinylate the C termini of membrane proteins, allowing these proteins to be tetramerized by binding to streptavidin, Because of the specificity of the biotinylation at the C terminus, the orientation of the membrane proteins on the streptavidin is equivalent to that of the native protein on the cell surface. These tetrameric proteins can be used to study protein receptor-ligand interactions at the cell surface, and site-specific biotinylation can be used to study proteins in vitro using a defined orientation. (C) 1999 Academic Press.
引用
收藏
页码:9 / 15
页数:7
相关论文
共 14 条
[1]   COOPERATIVE BINDING OF THE ESCHERICHIA-COLI REPRESSOR OF BIOTIN BIOSYNTHESIS TO THE BIOTIN OPERATOR SEQUENCE [J].
ABBOTT, J ;
BECKETT, D .
BIOCHEMISTRY, 1993, 32 (37) :9649-9656
[2]   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
[3]   HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C [J].
Braud, VM ;
Allan, DSJ ;
O'Callaghan, CA ;
Söderström, K ;
D'Andrea, A ;
Ogg, GS ;
Lazetic, S ;
Young, NT ;
Bell, JI ;
Phillips, JH ;
Lanier, LL ;
McMichael, AJ .
NATURE, 1998, 391 (6669) :795-799
[4]  
BUONCRISTIANI MR, 1988, J BIOL CHEM, V263, P1013
[5]   Direct visualization of antigen-specific CD8+ T cells during the primary immune response to Epstein-Barr virus in vivo [J].
Callan, MFC ;
Tan, L ;
Annels, N ;
Ogg, GS ;
Wilson, JDK ;
O'Callaghan, CA ;
Steven, N ;
McMichael, AJ ;
Rickinson, AB .
JOURNAL OF EXPERIMENTAL MEDICINE, 1998, 187 (09) :1395-1402
[6]   CONTROL OF TRANSLATION BY MESSENGER-RNA SECONDARY STRUCTURE IN ESCHERICHIA-COLI - A QUANTITATIVE-ANALYSIS OF LITERATURE DATA [J].
DESMIT, MH ;
VANDUIN, J .
JOURNAL OF MOLECULAR BIOLOGY, 1994, 244 (02) :144-150
[7]   A new look at T cells [J].
McMichael, AJ ;
O'Callaghan, CA .
JOURNAL OF EXPERIMENTAL MEDICINE, 1998, 187 (09) :1367-1371
[8]   Structural features impose tight peptide binding specificity in the nonclassical MHC molecule HLA-E [J].
O'Callaghan, CA ;
Tormo, J ;
Willcox, BE ;
Braud, VM ;
Jakobsen, BK ;
Stuart, DI ;
McMichael, AJ ;
Bell, JI ;
Jones, EY .
MOLECULAR CELL, 1998, 1 (04) :531-541
[9]   BIOTINYL 5'-ADENYLATE - COREPRESSOR ROLE IN THE REGULATION OF THE BIOTIN GENES OF ESCHERICHIA-COLI K-12 [J].
PRAKASH, O ;
EISENBERG, MA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (11) :5592-5595
[10]   Production and crystallization of MHC class I B allele single peptide complexes [J].
Reid, SW ;
Smith, KJ ;
Jakobsen, BK ;
OCallaghan, CA ;
Reyburn, H ;
Harlos, K ;
Stuart, DI ;
McMichael, AJ ;
Bell, JI ;
Jones, EY .
FEBS LETTERS, 1996, 383 (1-2) :119-123