Deciphering the function of an ORF:: Salmonella enterica DeoM protein is a new mutarotase specific for deoxyribose

被引:5
作者
Assairi, L
Bertrand, T
Ferdinand, J
Slavova-Azmanova, N
Christensen, M
Briozzo, P
Schaeffer, F
Craescu, CT
Neuhard, J
Bârzu, O
Gilles, AM
机构
[1] Inst Pasteur, Unite Genet Genom Bacteriens, F-75724 Paris 15, France
[2] Inst Pasteur, Lab Chim Struct Macromol, F-75724 Paris 15, France
[3] Inst Pasteur, CNRS, URA 2185, Unite Biochim Struct, F-75724 Paris 15, France
[4] CNRS, Lab Enzymol & Biochim Struct, UPR 9063, F-91198 Gif Sur Yvette, France
[5] Univ Copenhagen, Inst Mol Biol, Dept Biol Chem, Copenhagen K, Denmark
[6] Inst Natl Agron Paris Grignon, Chim Biol Lab, F-78850 Thiverval Grignon, France
[7] Inst Natl Sante & Rech Med, U350, F-91405 Orsay, France
[8] Ctr Univ Paris Sud, Inst Curie, F-91405 Orsay, France
关键词
Salmonella enterica; DeoM protein; deoxyribose mutarotase; site-directed mutagenesis; structural characterization;
D O I
10.1110/ps.03566004
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
We identified in Salmonella enterica serovar Typhi a cluster of four genes encoding a deoxyribokinase (DeoK), a putative permease (DeoP), a repressor (DeoQ), and an open reading frame encoding a 337 amino acid residues protein of unknown function. We show that the latter protein, called DeoM, is a hexamer whose synthesis is increased by a factor over 5 after induction with deoxyribose. The CD spectrum of the purified recombinant protein indicated a dominant contribution of P-type secondary structure and a small content of alpha-helix. Temperature and guanidinium hydrochloride induced denaturation of DeoM indicated that the hexamer dissociation and monomer unfolding are coupled processes. DeoM exhibits 12.5% and 15% sequence identity with galactose mutarotase from Lactococcits lactis and respectively Escherichia coli, which suggested that these three proteins share similar functions. Polarimetric experiments demonstrated that DeoM is a mutarotase with high specificity for deoxyribose. Site-directed mutagenesis of His183 in DeoM, corresponding to a catalytically active residue in GalM, yielded an almost inactive deoxyribose mutarotase. DeoM was crystallized and diffraction data collected for two crystal systems, confirmed its hexameric state. The possible role of the protein and of the entire gene cluster is discussed in connection with the energy metabolism of S. enterica under particular growth conditions.
引用
收藏
页码:1295 / 1303
页数:9
相关论文
共 36 条
[1]   Galactose mutarotase: Purification, characterization, and investigations of two important histidine residues [J].
Beebe, JA ;
Frey, PA .
BIOCHEMISTRY, 1998, 37 (42) :14989-14997
[2]   DEPENDENCE OF LACTOSE METABOLISM UPON MUTAROTASE ENCODED IN THE GAL OPERON IN ESCHERICHIA-COLI [J].
BOUFFARD, GG ;
RUDD, KE ;
ADHYA, SL .
JOURNAL OF MOLECULAR BIOLOGY, 1994, 244 (03) :269-278
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]  
Brunger AT, 1998, ACTA CRYSTALLOGR D, V54, P905, DOI 10.1107/s0907444998003254
[5]   Structural genomics: beyond the Human Genome Project [J].
Burley, SK ;
Almo, SC ;
Bonanno, JB ;
Capel, M ;
Chance, MR ;
Gaasterland, T ;
Lin, DW ;
Sali, A ;
Studier, FW ;
Swaminathan, S .
NATURE GENETICS, 1999, 23 (02) :151-157
[6]   Regulation of expression of the 2-DeOXY-D-ribose utilization regulon, deoQKPX, from Salmonella enterica serovar typhimurium [J].
Christensen, M ;
Borza, T ;
Dandanell, G ;
Gilles, AM ;
Barzu, O ;
Kelln, RA ;
Neuhard, J .
JOURNAL OF BACTERIOLOGY, 2003, 185 (20) :6042-6050
[7]   Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods [J].
delaFortelle, E ;
Bricogne, G .
MACROMOLECULAR CRYSTALLOGRAPHY, PT A, 1997, 276 :472-494
[8]   Genetic evidence for a defective xylan degradation pathway in Lactococcus lactis [J].
Erlandson, KA ;
Delamarre, SC ;
Batt, CA .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (04) :1445-1452
[9]   A NOVEL GENETIC SYSTEM TO DETECT PROTEIN PROTEIN INTERACTIONS [J].
FIELDS, S ;
SONG, OK .
NATURE, 1989, 340 (6230) :245-246
[10]   DNA as a nutrient novel role for bacterial competence gene homologs [J].
Finkel, SE ;
Kolter, R .
JOURNAL OF BACTERIOLOGY, 2001, 183 (21) :6288-6293