Incorporation of β-selenolo[3,2-b]pyrrolyl-alanine into proteins for phase determination in protein X-ray crystallography

被引:37
作者
Bae, JH
Alefelder, S
Kaiser, JT
Friedrich, R
Moroder, L
Huber, R
Budisa, N
机构
[1] Max Planck Inst Biochem, D-82152 Martinsried, Germany
[2] Proteros Biostruct GmbH, D-82152 Martinsried, Germany
关键词
analogue incorporation; MAD; X-ray crystallography; seleno-methionine; tryptophan;
D O I
10.1006/jmbi.2001.4699
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
beta -Selenolo[3,2-b]pyrrolyl-L-alanine that mimics tryptophan with the benzene ring of the indole moiety replaced by selenophene, was incorporated into human annexin V and barstar. This was achieved by fermentation and expression in a Trp-auxotrophic Escherichia coli host strain using the selective pressure incorporation method. The selenoproteins were obtained in yields comparable to those of the wild-type proteins and exhibit full crystallographic isomorphism to the parent proteins, but expectedly show altered absorbance profiles and quenched tryptophan fluorescence. Since the occurrence of tryptophan residues in proteins is rare, incorporation of the electron-rich selenium-containing tryptophan surrogate into proteins represents a useful supplementation and even a promising novel alternative to selenomethionine for solving the phase problem in protein X-ray crystallography. (C) 2001 Academic Press.
引用
收藏
页码:925 / 936
页数:12
相关论文
共 46 条
[1]   THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].
BAILEY, S .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763
[2]   STRUCTURE-FUNCTION ANALYSIS OF THE ION-CHANNEL SELECTIVITY FILTER IN HUMAN ANNEXIN-V [J].
BERENDES, R ;
VOGES, D ;
DEMANGE, P ;
HUBER, R ;
BURGER, A .
SCIENCE, 1993, 262 (5132) :427-430
[3]   Screening for phasing atoms in protein crystallography [J].
Boggon, TJ ;
Shapiro, L .
STRUCTURE, 2000, 8 (07) :R143-R149
[4]   Crystallography & NMR system:: A new software suite for macromolecular structure determination [J].
Brunger, AT ;
Adams, PD ;
Clore, GM ;
DeLano, WL ;
Gros, P ;
Grosse-Kunstleve, RW ;
Jiang, JS ;
Kuszewski, J ;
Nilges, M ;
Pannu, NS ;
Read, RJ ;
Rice, LM ;
Simonson, T ;
Warren, GL .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1998, 54 :905-921
[5]   Structure and evolution of the genetic code viewed from the perspective of the experimentally expanded amino acid repertoire in vivo [J].
Budisa, N ;
Moroder, L ;
Huber, R .
CELLULAR AND MOLECULAR LIFE SCIENCES, 1999, 55 (12) :1626-1635
[6]   Residue-specific bioincorporation of non-natural, biologically active amino acids into proteins as possible drug carriers:: Structure and stability of the per-thiaproline mutant of annexin V [J].
Budisa, N ;
Minks, C ;
Medrano, FJ ;
Lutz, J ;
Huber, R ;
Moroder, L .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (02) :455-459
[7]  
BUDISA N, 1995, EUR J BIOCHEM, V230, P788
[8]   Bioincorporation of telluromethionine into proteins: A promising new approach for X-ray structure analysis of proteins [J].
Budisa, N ;
Karnbrock, W ;
STeinbacher, S ;
Humm, A ;
Prade, L ;
Neuefeind, T ;
Moroder, L ;
Huber, R .
JOURNAL OF MOLECULAR BIOLOGY, 1997, 270 (04) :616-623
[9]   Toward the experimental codon reassignment in vivo:: protein building with an expanded amino acid repertoire [J].
Budisa, N ;
Minks, C ;
Alefelder, S ;
Wenger, W ;
Dong, FM ;
Moroder, L ;
Huber, R .
FASEB JOURNAL, 1999, 13 (01) :41-51
[10]  
BUDISA N, 2001, IN PRESS PROTEIN SCI