Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2003-5-14
pubmed:databankReference
pubmed:abstractText
N-acetylneuraminate lyase (NAL) and dihydrodipicolinate synthase (DHDPS) belong to the NAL subfamily of (betaalpha)(8)-barrels. They share a common catalytic step but catalyze reactions in different biological pathways. By rational design, we have introduced various mutations into the NAL scaffold from Escherichia coli to switch the activity toward DHDPS. These mutants were tested with respect to their catalytic properties in vivo and in vitro as well as their stability. One point mutation (L142R) was sufficient to create an enzyme that could complement a bacterial auxotroph lacking the gene for DHDPS as efficiently as DHDPS itself. In vitro, this mutant had an increased DHDPS activity of up to 19-fold as defined by the specificity constant k(cat)K(M) for the new substrate l-aspartate-beta-semialdehyde when compared with the residual activity of NAL wild-type, mainly because of an increased turnover rate. At the same time, mutant L142R maintained much of its original NAL activity. We have solved the crystal structure of mutant L142R at 1.8 A resolution in complex with the inhibitor beta-hydroxypyruvate. This structure reveals that the conformations of neighboring active site residues are left virtually unchanged by the mutation. The high flexibility of R142 may favor its role in assisting in catalysis. Perhaps, nature has exploited the catalytic promiscuity of many enzymes to evolve novel enzymes or biological pathways during the course of evolution.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-10099128, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-10758475, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-10944186, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-10968789, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11031117, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11054297, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11135667, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11196650, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11196654, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11196655, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11432751, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11518524, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-11950559, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-14353904, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-1458058, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-15299374, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-1673060, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-2025413, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-2610349, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-3131636, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-7737457, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-7853400, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-791073, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-7984417, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-8081752, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-8993314, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-9047371, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-9048556, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-9165072, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-9417939, http://linkedlifedata.com/resource/pubmed/commentcorrection/12711733-9757107
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0027-8424
pubmed:author
pubmed:issnType
Print
pubmed:day
13
pubmed:volume
100
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5694-9
pubmed:dateRevised
2010-9-14
pubmed:meshHeading
pubmed-meshheading:12711733-Amino Acid Sequence, pubmed-meshheading:12711733-Amino Acid Substitution, pubmed-meshheading:12711733-Cloning, Molecular, pubmed-meshheading:12711733-Consensus Sequence, pubmed-meshheading:12711733-Crystallography, X-Ray, pubmed-meshheading:12711733-Escherichia coli, pubmed-meshheading:12711733-Evolution, Molecular, pubmed-meshheading:12711733-Hydro-Lyases, pubmed-meshheading:12711733-Kinetics, pubmed-meshheading:12711733-Models, Biological, pubmed-meshheading:12711733-Models, Molecular, pubmed-meshheading:12711733-Molecular Sequence Data, pubmed-meshheading:12711733-Mutagenesis, Site-Directed, pubmed-meshheading:12711733-Oxo-Acid-Lyases, pubmed-meshheading:12711733-Protein Conformation, pubmed-meshheading:12711733-Recombinant Proteins, pubmed-meshheading:12711733-Sequence Alignment, pubmed-meshheading:12711733-Sequence Homology, Amino Acid, pubmed-meshheading:12711733-Thermodynamics
pubmed:year
2003
pubmed:articleTitle
Mimicking natural evolution in vitro: an N-acetylneuraminate lyase mutant with an increased dihydrodipicolinate synthase activity.
pubmed:affiliation
Cambridge University Chemical Laboratory and Cambridge Centre for Protein Engineering, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH, United Kingdom.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't