Thermal unfolding of dodecameric glutamine synthetase: inhibition of aggregation by urea

Protein Science : a Publication of the Protein Society
N J Nosworthy, A Ginsburg

Abstract

Thermal unfolding of dodecameric manganese glutamine synthetase (622,000 M(r)) at pH 7 and approximately 0.02 ionic strength occurs in two observable steps: a small reversible transition (Tm approximately 42 degrees C; delta H approximately equal to 0.9 J/g) followed by a large irreversible transition (Tm approximately 81 degrees C; delta H approximately equal to 23.4 J/g) in which secondary structure is lost and soluble aggregates form. Secondary structure, hydrophobicity, and oligomeric structure of the equilibrium intermediate are the same as for the native protein, whereas some aromatic residues are more exposed. Urea (3 M) destabilizes the dodecamer (with a tertiary structure similar to that without urea at 55 degrees C) and inhibits aggregation accompanying unfolding at < or = 0.2 mg protein/mL. With increasing temperature (30-70 degrees C) or incubation times at 25 degrees C (5-35 h) in 3 M urea, only dodecamer and unfolded monomer are detected. In addition, the loss in enzyme secondary structure is pseudo-first-order (t1/2 = 1,030 s at 20.0 degrees C in 4.5 M urea). Differential scanning calorimetry of the enzyme in 3 M urea shows one endotherm (Tmax approximately 64 degrees C; delta H = 17 +/- 2 J/g). The enthalpy chan...Continue Reading

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Citations

Nov 29, 2007·Cell Biochemistry and Biophysics·Veronica PiercePaul E Smith
Mar 14, 2008·Assay and Drug Development Technologies·Peggy A ThompsonJames R Appleman
Nov 10, 2009·The Journal of Chemical Physics·Moon Bae Gee, Paul E Smith
Mar 12, 2016·The Journal of Physical Chemistry. B·Gouri S JasKrzysztof Kuczera
Jul 1, 2011·Molecular BioSystems·Jian-Yi PanXuan-Xian Peng
Aug 5, 2014·Molecular Biology Reports·Wei ZhaoQuan-Hong Yao
Dec 29, 2004·Biophysical Chemistry·Paul E Smith

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