Domains III and I-2{alpha}, at the entrance of the binding cleft, play an important role in cold adaptation of the periplasmic dipeptide-binding protein (DppA) from the deep-sea psychrophilic bacterium Pseudoalteromonas sp. strain SM9913.
The peptide transporter from a cold-adapted bacterium has never been reported. In the present study, the dpp operon from the psychrophilic bacterium Pseudoalteromonas sp. strain SM9913 was cloned and analyzed. The dipeptide binding protein DppA of SM9913 was overexpressed in Escherichia coli, and its cold adaptation characteristics were studied. The recombinant DppA of SM9913 (PsDppA) displayed the highest ligand-binding affinity at 15 degrees C, whereas the recombinant DppA of E. coli (EcDppA) displayed the highest ligand-binding affinity at 35 degrees C. Thermal and guanidium hydrochloride unfolding analyses indicated that PsDppA has more structural instability than EcDppA. Six domain-exchanged mutants of PsDppA were expressed and purified. Analyses of these mutants indicated that domains III, I-2, and I-3 of PsDppA were less stable than those from EcDppA and that domains III and I-2 made a significant contribution to the high binding affinity of PsDppA at low temperatures. Structural and sequence analyses suggested that the state transition-involved regions in domain III and the alpha part of domain I-2 are the hot spots of optimization during cold adaptation and that decreasing the side-chain size in these regions is an important strategy for the cold adaptation of PsDppA.
Zhang
Wei-Xin
WX
State Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Shandong University, Jinan 250100, People's Republic of China.
Xie
Bin-Bin
BB
Chen
Xiu-Lan
XL
Dong
Sheng
S
Zhang
Xi-Ying
XY
Zhou
Bai-Cheng
BC
Zhang
Yu-Zhong
YZ
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D003080:Cold Temperature; D004151:Dipeptides / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D033902:Periplasmic Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D031161:Pseudoalteromonas / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012623:Seawater / Q000382:microbiology*; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D004151:Dipeptides; D029968:Escherichia coli Proteins; D033902:Periplasmic Binding Proteins; C066768:dppA protein, E coli
J Biol Chem. 1985 Sep 5;260(19):10812-8
3897225
Biochemistry. 2000 Apr 25;39(16):4855-62
10769143
Structure. 1995 Dec 15;3(12):1395-406
8747465
Annu Rev Biochem. 2006;75:403-33
16756497
J Bacteriol. 1984 Oct;160(1):122-30
6090406
Nat Rev Microbiol. 2003 Dec;1(3):200-8
15035024
J Bacteriol. 1991 Jan;173(1):234-44
1702779
J Bacteriol. 1983 Sep;155(3):1434-8
6350270
EMBO J. 2009 May 6;28(9):1332-40
19300437
Science. 1994 Jun 10;264(5165):1578-81
8202710
J Biol Chem. 1982 Feb 10;257(3):1131-3
7035444
J Biol Chem. 1997 Jun 20;272(25):15661-7
9188456
Genome Res. 2005 Oct;15(10):1325-35
16169927
J Bacteriol. 2004 Jan;186(1):51-60
14679224
Cell Mol Life Sci. 2003 Oct;60(10):2100-14
14618258
Microbiology (Reading). 2007 Jul;153(Pt 7):2116-2125
17600056
Biochemistry. 1999 Nov 2;38(44):14440-50
10545166
Biochemistry. 1995 Dec 26;34(51):16585-95
8527431
J Mol Biol. 1993 Dec 5;234(3):779-815
8254673
Science. 1997 Sep 5;277(5331):1453-62
9278503
J Biol Chem. 2008 Dec 26;283(52):36100-7
18977758
Naturwissenschaften. 2007 Feb;94(2):77-99
17039344
EMBO J. 2000 Jul 17;19(14):3649-56
10899119
Protein Sci. 1995 Nov;4(11):2327-34
8563629
Microbiology (Reading). 1999 Oct;145 ( Pt 10):2891-901
10537211
FEBS J. 2008 Jul;275(13):3290-8
18485005
J Bacteriol. 1986 Nov;168(2):775-9
3536860
Microbiology (Reading). 2007 May;153(Pt 5):1566-1572
17464071
Proc Natl Acad Sci U S A. 1999 May 25;96(11):6161-5
10339558
Adv Microb Physiol. 1994;36:1-80
7942312