|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
32
|
|
pubmed:dateCreated |
2005-8-10
|
|
pubmed:abstractText |
We describe an iterative approach for creating protein switches involving the in vitro recombination of two nonhomologous genes. We demonstrate this approach by recombining the genes coding for TEM1 beta-lactamase (BLA) and the Escherichia coli maltose binding protein (MBP) to create a family of MBP-BLA hybrids in which maltose is a positive or negative effector of beta-lactam hydrolysis. Some of these MBP-BLA switches were effectively "on-off" in nature, with maltose altering catalytic activity by as much as 600-fold. The ability of these switches to confer an effector-dependent growth/no growth phenotype to E. coli cells was exploited to rapidly identify, from a library of 4 x 10(6) variants, MBP-BLA switch variants that respond to sucrose as the effector. The transplantation of these mutations into wild-type MBP converted MBP into a "sucrose-binding protein," illustrating the switches potential as a tool to rapidly identify ligand-binding proteins.
|
|
pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-10436083,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-10486551,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-10500161,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11013401,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11035796,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11320244,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11399072,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11504940,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11689849,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11875520,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-11914727,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-12082164,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-12204690,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-12483203,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-12736688,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-12963365,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-13897943,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-1436034,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-14512628,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-14527313,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-14581486,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-14717614,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-14741221,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15159530,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15247421,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15326589,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15382234,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15459282,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15555998,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15632292,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15657118,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-15734643,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-1930157,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-2211591,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-2744488,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-817286,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-8230196,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-9211908,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-9211910,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061816-9360947
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Aug
|
|
pubmed:issn |
0027-8424
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:day |
9
|
|
pubmed:volume |
102
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
11224-9
|
|
pubmed:dateRevised |
2010-11-18
|
|
pubmed:meshHeading |
pubmed-meshheading:16061816-Allosteric Regulation,
pubmed-meshheading:16061816-Carrier Proteins,
pubmed-meshheading:16061816-Catalysis,
pubmed-meshheading:16061816-Directed Molecular Evolution,
pubmed-meshheading:16061816-Escherichia coli,
pubmed-meshheading:16061816-Escherichia coli Proteins,
pubmed-meshheading:16061816-Genes, Switch,
pubmed-meshheading:16061816-Ligands,
pubmed-meshheading:16061816-Maltose-Binding Proteins,
pubmed-meshheading:16061816-Protein Binding,
pubmed-meshheading:16061816-Protein Engineering,
pubmed-meshheading:16061816-Sucrose,
pubmed-meshheading:16061816-beta-Lactamases
|
|
pubmed:year |
2005
|
|
pubmed:articleTitle |
Directed evolution of protein switches and their application to the creation of ligand-binding proteins.
|
|
pubmed:affiliation |
Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.
|
|
pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, N.I.H., Extramural
|
