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Re‐engineering a β‐lactamase using prototype peptides from a library of local structural motifs
Author(s) -
Risso Valeria A.,
Primo María E.,
Ermácora Mario R.
Publication year - 2009
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.47
Subject(s) - structural similarity , protein engineering , structural motif , computational biology , folding (dsp implementation) , sequence (biology) , protein folding , sequence space , protein structure , protein superfamily , protein secondary structure , recombinant dna , peptide sequence , protein design , biology , chemistry , stereochemistry , genetics , biochemistry , enzyme , mathematics , engineering , pure mathematics , banach space , gene , electrical engineering
B. licheniformis exo‐small β‐lactamase (ESBL) has a complex architecture with twelve α helices and a five‐stranded beta sheet. We replaced, separately or simultaneously, three of the ESBL α helices with prototype amphiphatic helices from a catalog of secondary structure elements. Although the substitutes bear no sequence similarity to the originals and pertain to unrelated protein families, all the engineered ESBL variants were found able to fold in native like structures with in vitro and in vivo enzymic activity. The triple substituted variant resembles a primitive protein, with folding defects such as a strong tendency to oligomerization and very low stability; however it mimics a non homologous recombinant abandoning the family sequence space while preserving fold. The results test protein folding and evolution theories.