Signalling substitutions in the periplasmic domain of chemoreceptor Trg induce or reduce helical sliding in the transmembrane domain

被引:12
作者
Beel, BD [1 ]
Hazelbauer, GL [1 ]
机构
[1] Washington State Univ, Sch Mol Biosci, Pullman, WA 99164 USA
关键词
D O I
10.1046/j.1365-2958.2001.02446.x
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 used in vivo oxidative cross-linking of engineered cysteine pairs to assess conformational changes in the four-helix transmembrane domain of chemoreceptor Trg. Extending previous work, we searched for and found a fourth cross-linking pair that spanned the intrasubunit interface between transmembrane helix 1 (TM1) and its partner TM2. We determined the effects of ligand occupancy on cross-linking rate constants for all four TM1-TM2 diagnostic pairs in conditions that allowed the formation of receptor-kinase complexes for the entire cellular complement of Trg. Occupancy altered all four rates in a pattern that implicated sliding of TM2 relative to TM1 towards the cytoplasm as the transmembrane signalling movement in receptor-kinase complexes. Transmembrane signalling can be reduced or induced by single amino acid substitutions in the ligand-binding region of the periplasmic domain of Trg. We determined the effects of these substitutions on conformation in the transmembrane domain and on ligand-induced changes using the diagnostic TM1TM2 cysteine pairs. Effects on rates of in vivo crosslinking showed that induced signalling substitutions altered the relative positions of TM1 and TM2 in the same way as ligand binding, and reduced signalling substitutions blocked or attenuated the ligand-induced shift. These results provide strong support for the helical sliding model of transmembrane signalling.
引用
收藏
页码:824 / 834
页数:11
相关论文
共 47 条
[1]   Comparison in vitro of a high- and a low-abundance chemoreceptor of Escherichia coli:: Similar kinase activation but different methyl-accepting activities [J].
Barnakov, AN ;
Barnakova, LA ;
Hazelbauer, GL .
JOURNAL OF BACTERIOLOGY, 1998, 180 (24) :6713-6718
[2]   Signaling domain of the aspartate receptor is a helical hairpin with a localized kinase docking surface: Cysteine and disulfide scanning studies [J].
Bass, RB ;
Coleman, MD ;
Falke, JJ .
BIOCHEMISTRY, 1999, 38 (29) :9317-9327
[3]   Substitutions in the periplasmic domain of low-abundance chemoreceptor Trg that induce or reduce transmembrane signaling: Kinase activation and context effects [J].
Beel, BD ;
Hazelbauer, GL .
JOURNAL OF BACTERIOLOGY, 2001, 183 (02) :671-679
[4]   An aspartate insulin receptor chimera mitogenically activates fibroblasts [J].
Biemann, HP ;
Harmer, SL ;
Koshland, DE .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (44) :27927-27930
[5]   THE 3-DIMENSIONAL STRUCTURE OF THE ASPARTATE RECEPTOR FROM ESCHERICHIA-COLI [J].
BOWIE, JU ;
PAKULA, AA ;
SIMON, MI .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1995, 51 :145-154
[6]  
BURROWS GG, 1989, J BIOL CHEM, V264, P17309
[7]   Cysteine and disulfide scanning reveals two amphiphilic helices in the linker region of the aspartate chemoreceptor [J].
Butler, SL ;
Falke, JJ .
BIOCHEMISTRY, 1998, 37 (30) :10746-10756
[8]   THERMAL MOTIONS OF SURFACE ALPHA-HELICES IN THE D-GALACTOSE CHEMOSENSORY RECEPTOR - DETECTION BY DISULFIDE TRAPPING [J].
CAREAGA, CL ;
FALKE, JJ .
JOURNAL OF MOLECULAR BIOLOGY, 1992, 226 (04) :1219-1235
[9]   LOCK ON OFF DISULFIDES IDENTIFY THE TRANSMEMBRANE SIGNALING HELIX OF THE ASPARTATE RECEPTOR [J].
CHERVITZ, SA ;
FALKE, JJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (41) :24043-24053
[10]   TRANSMEMBRANE SIGNALING BY THE ASPARTATE RECEPTOR - ENGINEERED DISULFIDES REVEAL STATIC REGIONS OF THE SUBUNIT INTERFACE [J].
CHERVITZ, SA ;
LIN, CM ;
FALKE, JJ .
BIOCHEMISTRY, 1995, 34 (30) :9722-9733