Stirring rate affects thermodynamics and unfolding kinetics in isothermal titration calorimetry.

Journal of Biochemistry
Takahiro MarunoSusumu Uchiyama

Abstract

Isothermal titration calorimetry (ITC) directly provides thermodynamic parameters depicting the energetics of intermolecular interactions in solution. During ITC experiments, a titration syringe with a paddle is continuously rotating to promote a homogeneous mixing. Here, we clarified that the shape of the paddles (flat, corkscrew and small-pitched corkscrew) and the stirring rates influence on the thermodynamic parameters of protein-ligand interaction. Stirring with the flat paddle at lower and higher rate both yielded a lower exothermic heat due to different reasons. The complete reaction with no incompetent fractions was achieved only when the stirring was performed at 500 or 750 rpm using the small-pitched corkscrew paddle. The evaluation of the protein solution after 1,500 rpm stirring indicated that proteins in the soluble fraction decreased to 94% of the initial amount, among which 6% was at an unfolded state. In addition, a significant increase of micron aggregates was confirmed. Furthermore, a new approach for the determination of the unfolding kinetics based on the time dependence of the total reaction heat was developed. This study demonstrates that a proper stirring rate and paddle shape are essential for the reliab...Continue Reading

References

Jan 1, 1979·Advances in Protein Chemistry·P L Privalov
Jan 1, 1990·Critical Reviews in Biochemistry and Molecular Biology·P L Privalov
Jul 25, 1986·Biochimica Et Biophysica Acta·W Pfeil, P I Ohlsson
Aug 31, 1999·Nature Structural Biology·T M RaschkeS Marqusee
Jan 18, 2003·Chemistry : a European Journal·Siegfried Höfinger, Francesco Zerbetto
Dec 4, 2003·Journal of the American Chemical Society·W Bruce Turnbull, Antonio H Daranas
Oct 20, 2005·Journal of Bioscience and Bioengineering·K NakanishiK Imamura
Oct 19, 2006·Biochimica Et Biophysica Acta·Tsutomu ArakawaKouhei Tsumoto
Nov 23, 2007·Biochemical Society Transactions·D J Brockwell
Apr 23, 2008·Current Protocols in Protein Science·Gerald R Grimsley, C Nick Pace
Aug 20, 1996·Biotechnology and Bioengineering·Y F Maa, C C Hsu
Jun 20, 1997·Biotechnology and Bioengineering·Y F Maa, C C Hsu
Apr 17, 2009·Biotechnology and Bioengineering·Jared S BeeTheodore W Randolph
May 20, 2009·Biophysical Journal·Lorna AshtonEwan W Blanch
Jun 18, 2009·Combinatorial Chemistry & High Throughput Screening·William B PetersRichard K Brown
Sep 3, 2009·Biotechnology and Bioengineering·Jared S BeeTheodore W Randolph
May 6, 2010·Biophysical Journal·Patrick J B EdwardsMartin A K Williams
Feb 8, 2011·Advances in Colloid and Interface Science·Michael RabeStefan Seeger
Mar 30, 2011·Analytical Biochemistry·Joel Tellinghuisen, John D Chodera
Apr 2, 2011·The Journal of Biological Chemistry·Marisa K JoubertLinda O Narhi
Aug 23, 2011·Analytical Biochemistry·Jana BroeckerSandro Keller
Apr 26, 2012·Analytical Chemistry·Sandro KellerPeter Schuck
Dec 1, 2012·MAbs·Ruairidh G CoustonChristopher F van der Walle
Jun 12, 2013·Colloids and Surfaces. B, Biointerfaces·Yang WeiRobert A Latour
Nov 25, 2014·Journal of Pharmaceutical Sciences·Theodore W RandolphCorrine S Lengsfeld
Nov 25, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Yang WeiRobert A Latour
Feb 18, 2015·Scientific Reports·Shusaku HaradaTsuyoshi Nakanishi
Feb 13, 2016·Journal of Pharmaceutical Sciences·Ahmad S SediqM Reza Nejadnik
May 2, 2016·Journal of Pharmaceutical Sciences·Lukas BrücklCorinna Sonderegger
Sep 7, 2016·Biochimica Et Biophysica Acta. General Subjects·Samuel A KantonenMichael K Gilson
Nov 27, 2016·Journal of Pharmaceutical Sciences·Tetsuo TorisuSusumu Uchiyama
Jun 13, 2017·Journal of Pharmaceutical Sciences·Tetsuo TorisuSusumu Uchiyama
Feb 9, 2018·Journal of Pharmaceutical Sciences·Takahiro MarunoSusumu Uchiyama
Feb 27, 2018·Analytical Chemistry·Robert EvansGareth A Morris
Apr 4, 2018·Engineering in Life Sciences·Mark DuerkopAlois Jungbauer

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