Loop size optimization induces a strong thermal stabilization of the thioredoxin fold(137 views visite) Ruggiero A, Smaldone G, Esposito L, Balasco N, Vitagliano L
Keywords Parole chiave: Loop Size Optimization, Protein Engineering, Protein Thermal Stability, Protocol Of Protein Stabilization, Thioredoxin Chimeric Proteins, Amino Acid Sequence
, Crystallography, X-Ray
, Escherichia Coli Chemistry
, Humans
, Models, Molecular
, Protein Folding
, Protein Stability
, Sequence Alignment
, Sulfolobus Chemistry
, Sulfolobus Solfataricus Chemistry
, Temperature
, Thioredoxins Chemistry Isolation, Purification Metabolism
Affiliations Affiliazioni: *** IBB - CNR ***
Institute of Biostructures and Bioimaging, C.N.R., Naples, Italy., IRCCS SDN, Naples, Italy.,
IRCCS SDN, Naples, Italy.
References Riferimenti: Not available. Non disponibili.
Loop size optimization induces a strong thermal stabilization of the thioredoxin fold
The definition of the structural basis of protein thermostability represents a major topic in structural biology and protein chemistry. We have recently observed that proteins isolated from thermophilic organisms show a better adherence to the fundamental rules of protein topology previously unveiled by Baker and coworkers (Koga et al. Nature. 2012; 491: 222-227). Here, we explored the possibility that ad hoc modifications of a natural protein following these rules could represent an efficient tool to stabilize its structure. Hence, we here designed and characterized novel variants of Escherichia coli thioredoxin (EcTrx) using a repertoire of biophysical/structural techniques. Trx chimeric variants were prepared by replacing the loop of EcTrx with the corresponding ones present in the Trxs isolated from Sulfolobus solfataricus and Sulfolobus tokodaii that show a better adherence to the topological rules. Interestingly, although the loop sequences of these proteins did not display any significant similarity, their insertion in EcTrx induced a remarkable stabilization of the protein (>/=10 degrees C). The crystallographic structure of one of these variants corroborates the hypothesis that the optimization of the loop size is the driving force of the observed stabilization. The remarkable stabilization of the two novel chimeric Trxs, generated by applying the topological rules, represents the proof of concept that these rules may be used to stabilize natural proteins through the ad hoc optimization of the loop size. Based on the present results, we propose a novel protocol of protein stabilization that can be potentially applied to other proteins.
Loop size optimization induces a strong thermal stabilization of the thioredoxin fold
Aloj L, Aurilio M, Rinaldi V, D'Ambrosio L, Tesauro D, Peitl PK, Maina T, Mansi R, Von Guggenberg E, Joosten L, Sosabowski JK, Breeman WA, De Blois E, Koelewijn S, Melis M, Waser B, Beetschen K, Reubi JC, De Jong M * The EEE project(387 visite) Proc Int Cosm Ray Conf Icrc Universidad Nacional Autonoma De Mexico, 2007; 5(HEPART2): 977-980. Impact Factor:0 DettagliEsporta in BibTeXEsporta in EndNote
561 Records (552 escludendo Abstract e Conferenze). Impact factor totale: 2189.447 (2162.267 escludendo Abstract e Conferenze). Impact factor a 5 anni totale: 2272.469 (2241.432 escludendo Abstract e Conferenze).
Last modified by Ultima modifica di Marco Comerci on in data Wednesday 02 December 2020, 18:09:27 137 views visite. Last view on Ultima visita in data Friday 26 February 2021, 8:27:42