| RRC ID |
53671
|
| 著者 |
Sahoo A, Khare S, Devanarayanan S, Jain PC, Varadarajan R.
|
| タイトル |
Residue proximity information and protein model discrimination using saturation-suppressor mutagenesis.
|
| ジャーナル |
Elife
|
| Abstract |
Identification of residue-residue contacts from primary sequence can be used to guide protein structure prediction. Using Escherichia coli CcdB as the test case, we describe an experimental method termed saturation-suppressor mutagenesis to acquire residue contact information. In this methodology, for each of five inactive CcdB mutants, exhaustive screens for suppressors were performed. Proximal suppressors were accurately discriminated from distal suppressors based on their phenotypes when present as single mutants. Experimentally identified putative proximal pairs formed spatial constraints to recover >98% of native-like models of CcdB from a decoy dataset. Suppressor methodology was also applied to the integral membrane protein, diacylglycerol kinase A where the structures determined by X-ray crystallography and NMR were significantly different. Suppressor as well as sequence co-variation data clearly point to the X-ray structure being the functional one adopted in vivo. The methodology is applicable to any macromolecular system for which a convenient phenotypic assay exists.
|
| 巻・号 |
4
|
| 公開日 |
2015-12-30
|
| DOI |
10.7554/eLife.09532
|
| PII |
e09532
|
| PMID |
26716404
|
| PMC |
PMC4758949
|
| MeSH |
Bacterial Proteins / chemistry*
Bacterial Proteins / genetics
Diacylglycerol Kinase / chemistry*
Diacylglycerol Kinase / genetics
Escherichia coli / chemistry
Escherichia coli / enzymology*
Escherichia coli / genetics
Models, Molecular
Mutagenesis*
Protein Conformation
Suppression, Genetic*
|
| IF |
7.08
|
| 引用数 |
13
|
| リソース情報 |
| 原核生物(大腸菌) |
|
