Prediction and experimental validation of enzyme substrate specificity in protein structures.
Proc Natl Acad Sci U S A
; 110(45): E4195-202, 2013 Nov 05.
Article
em En
| MEDLINE
| ID: mdl-24145433
ABSTRACT
Structural Genomics aims to elucidate protein structures to identify their functions. Unfortunately, the variation of just a few residues can be enough to alter activity or binding specificity and limit the functional resolution of annotations based on sequence and structure; in enzymes, substrates are especially difficult to predict. Here, large-scale controls and direct experiments show that the local similarity of five or six residues selected because they are evolutionarily important and on the protein surface can suffice to identify an enzyme activity and substrate. A motif of five residues predicted that a previously uncharacterized Silicibacter sp. protein was a carboxylesterase for short fatty acyl chains, similar to hormone-sensitive-lipase-like proteins that share less than 20% sequence identity. Assays and directed mutations confirmed this activity and showed that the motif was essential for catalysis and substrate specificity. We conclude that evolutionary and structural information may be combined on a Structural Genomics scale to create motifs of mixed catalytic and noncatalytic residues that identify enzyme activity and substrate specificity.
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Bases de dados:
MEDLINE
Assunto principal:
Biologia Computacional
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Proteômica
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Enzimas
Tipo de estudo:
Evaluation_studies
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Prognostic_studies
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Risk_factors_studies
Idioma:
En
Revista:
Proc Natl Acad Sci U S A
Ano de publicação:
2013
Tipo de documento:
Article