| RRC ID |
3282
|
| 著者 |
Loftus SR, Walker D, Maté MJ, Bonsor DA, James R, Moore GR, Kleanthous C.
|
| タイトル |
Competitive recruitment of the periplasmic translocation portal TolB by a natively disordered domain of colicin E9.
|
| ジャーナル |
Proc Natl Acad Sci U S A
|
| Abstract |
The natively disordered N-terminal 83-aa translocation (T) domain of E group nuclease colicins recruits OmpF to a colicin-receptor complex in the outer membrane (OM) as well as TolB in the periplasm of Escherichia coli, the latter triggering translocation of the toxin across the OM. We have identified the 16-residue TolB binding epitope in the natively disordered T-domain of the nuclease colicin E9 (ColE9) and solved the crystal structure of the complex. ColE9 folds into a distorted hairpin within a canyon of the six-bladed beta-propeller of TolB, using two tryptophans to bolt the toxin to the canyon floor and numerous intramolecular hydrogen bonds to stabilize the bound conformation. This mode of binding enables colicin side chains to hydrogen-bond TolB residues in and around the channel that runs through the beta-propeller and that constitutes the binding site of peptidoglycan-associated lipoprotein (Pal). Pal is a globular binding partner of TolB, and their association is known to be important for OM integrity. The structure is therefore consistent with translocation models wherein the colicin disrupts the TolB-Pal complex causing local instability of the OM as a prelude to toxin import. Intriguingly, Ca(2+) ions, which bind within the beta-propeller channel and switch the surface electrostatics from negative to positive, are needed for the negatively charged T-domain to bind TolB with an affinity equivalent to that of Pal and competitively displace it. Our study demonstrates that natively disordered proteins can compete with globular proteins for binding to folded scaffolds but that this can require cofactors such as metal ions to offset unfavorable interactions.
|
| 巻・号 |
103(33)
|
| ページ |
12353-8
|
| 公開日 |
2006-8-15
|
| DOI |
10.1073/pnas.0603433103
|
| PII |
0603433103
|
| PMID |
16894158
|
| PMC |
PMC1567883
|
| MeSH |
Binding Sites
Calcium / metabolism
Colicins / chemistry*
Colicins / genetics
Colicins / metabolism*
Crystallography, X-Ray
Escherichia coli Proteins / chemistry*
Escherichia coli Proteins / genetics
Escherichia coli Proteins / metabolism*
Hydrogen Bonding
Models, Molecular
Molecular Sequence Data
Periplasmic Proteins / chemistry*
Periplasmic Proteins / genetics
Periplasmic Proteins / metabolism*
Protein Structure, Tertiary*
Thermodynamics
|
| IF |
9.412
|
| 引用数 |
52
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 原核生物(大腸菌) |
JW5110(BW25113 delta tolB) |
