PMID: 11342714May 9, 2001Paper

Stabilization of apoflavodoxin by replacing hydrogen-bonded charged Asp or Glu residues by the neutral isosteric Asn or Gln

Protein Engineering
María P IrúnJ Sancho

Abstract

Knowledge of protein stability principles provides a means to increase protein stability in a rational way. Here we explore the feasibility of stabilizing proteins by replacing solvent-exposed hydrogen-bonded charged Asp or Glu residues by the neutral isosteric Asn or GLN: The rationale behind this is a previous observation that, in some cases, neutral hydrogen bonds may be more stable that charged ones. We identified, in the apoflavodoxin from Anabaena PCC 7119, three surface-exposed aspartate or glutamate residues involved in hydrogen bonding with a single partner and we mutated them to asparagine or glutamine, respectively. The effect of the mutations on apoflavodoxin stability was measured by both urea and temperature denaturation. We observed that the three mutant proteins are more stable than wild-type (on average 0.43 kcal/mol from urea denaturation and 2.8 degrees C from a two-state analysis of fluorescence thermal unfolding data). At high ionic strength, where potential electrostatic repulsions in the acidic apoflavodoxin should be masked, the three mutants are similarly more stable (on average 0.46 kcal/mol). To rule out further that the stabilization observed is due to removal of electrostatic repulsions in apoflavod...Continue Reading

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Citations

Oct 19, 2005·Journal of Computer-aided Molecular Design·Claudia MachicadoJavier Sancho
May 9, 2012·Applied Biochemistry and Biotechnology·Debamitra ChakravortySanjukta Patra
Apr 23, 2002·Protein Science : a Publication of the Protein Society·Susana MaldonadoJavier Sancho
Jul 4, 2008·The Journal of Clinical Investigation·Angel L PeyAurora Martinez
Aug 7, 2002·Bioscience, Biotechnology, and Biochemistry·Shin-ichi SakasegawaToshihisa Ohshima
May 12, 2010·Journal of Structural Biology·Sara Ayuso-TejedorJavier Sancho
Oct 31, 2015·Applied Biochemistry and Biotechnology·Fucheng ZhuBingfang He
Mar 17, 2015·Scientific Reports·Emilio LamazaresJavier Sancho
Aug 1, 2006·Protein Science : a Publication of the Protein Society·Marta BuenoJavier Sancho
Feb 1, 2005·Biochimica Et Biophysica Acta·Ulrich Kragh-HansenMasaki Otagiri
Mar 28, 2006·Journal of Molecular Biology·Marta BuenoJavier Sancho
Dec 21, 2005·Journal of Molecular Biology·J López-LlanoJ Sancho
Jun 18, 2010·Journal of Molecular Biology·Sara Ayuso-TejedorM Angeles Jiménez
Aug 16, 2011·Biophysical Chemistry·Salvador Casares-AtienzaMaria M Garcia-Mira
Sep 6, 2000·Journal of Molecular Biology·R WintjensE Buisine
Mar 26, 2004·Biophysical Journal·Raul Perez-JimenezJose M Sanchez-Ruiz
Jan 18, 2017·Journal of Theoretical Biology·Mansour PoorebrahimHamzeh Rahimi
Aug 20, 2004·The Journal of Biological Chemistry·Jon López-LlanoJavier Sancho
Nov 14, 2007·The Journal of Biological Chemistry·Nunilo CremadesJavier Sancho
Jun 30, 2005·Journal of the American Chemical Society·Ivana Nikolic-HughesDaniel Herschlag

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