Unfolded, oxidized, and thermoinactivated forms of glyceraldehyde-3-phosphate dehydrogenase interact with the chaperonin GroEL in different ways

Oxidation of an Exposed Methionine Instigates the Aggregation of Glyceraldehyde-3-phosphate Dehydrogenase

“…There are numerous reports investigating the mechanism of GAPDH aggregation (11,13,22,23,(43)(44)(45)(46)(47) with several studies stipulating an essential role for intermolecular disulfide bond formation (11,13,22,23). These models of GAPDH aggregation are supported by the observation that insoluble disulfide-cross-linked forms of GAPDH have been found in vivo (11)(12)(13)18).…”

Studies on the Thermodenaturation Behavior of Bacillus subtilisα‐Amylase on Chromatographic Media

“…For the intrinsic fluorescence emission spectra, take the above Bacillus subtilis α-amylase solutions and assay their intrinsic fluorescence emission spectra on a Hitachi F-4500 fluorescence spectrometer. The excitation wavelength was 295 nm, both the excitation and emission slits were 10 nm, the scanning voltage and scanning rate were separately 700 mV and 1200 nm/s, the scanning wavelength area was 300-420 nm, and the experimental temperature was 25 . ℃…”

“…℃ Bacillus subtilis α-amylase concentration was 30 µg/mL. Fluorescence was excited at 295 nm and measurements were carried out at 25 . ℃…”

“…[2][3][4][5][6][7][8][9][10][11][12][13][14] Since the medium-term of the eighties of the 20th century, studies on the thermal behaviors of solute molecules have been chiefly focused on small solute molecules in reversed-phase liquid chromatography, and these involved in protein molecules have been rarely reported. [15][16][17][18][19] Karger et al 20 studied the thermal behaviors of lysozyme, α-lactalbumin and β-globulin on a hydrophobic ether bonded-phase medium by using parameter Z (the slope of plot of lg k' to ln B, k' is the capacity factor and B is the volume fraction of organic moderator in mobile phase); cooperating with other authors, one of the authors in this paper also studied the thermal behaviors of cytochrome C, myoglobin, bovine serum albumin, lysozyme, α-amylase and insulin on a PEG-400 hydrophobic medium and determined their thermodynamic parameters; 21 Marion and Peter 22 studied the heat-induced denaturation and aggregation of β-lactoglobulin as a function of pH in the range 6.0-8.0; Andrew and Jeffrey 23 studied the effect of pH on thermal denaturation of four main whey protein fractions in skim milk by gel permeation fast protein liquid chromatography (FPLC); Antonucci et al 24 studied the minor conformational changes of a doxorubicin-peptide conjugate; Naletova et al 25 studied the interaction of GroEL with different denatured forms of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and found that GroEL did not prevent thermal denaturation of GAPDH, but effectively interacted with the thermodenatured enzyme, thus preventing the aggregation of denatured molecules.…”

Thermodynamics and Extra‐thermodynamics of Bacillus subtilis α‐Amylase in Some Chromatographic Systems

“…[1][2][3][4][5] As for the protein molecules with a more complex structure, so far, the studies on their thermal behavior and thermodynamics on chromatographic media are rarely reported for their structures can be affected not only by the experimental temperature but also by the chromatographic media. Karger et al 6 studied the thermal stability of lysozyme, α-lactalbumin and β-globulin on a hydrophobic ether bonded-phase medium by using parameter Z, Esquibel-King et al 7 determined the enthalpy change and entropy change of bovine serum albumin on a hydrophobic chromatographic medium by using the non-linear Van't Hoff plot; Horvath et al 8 studied the thermodynamic behavior of some amino acid derivants on a hydrophobic chromatographic medium; co-operating with other authors, one of the authors in this paper also studied the thermal behavior of cytochrome c, myoglobin, bovine serum albumin, lysozyme, α-amylase and insulin on a PEG-400 hydrophobic medium and determined their thermodynamic parameters; 9 Hoffmann et al 10 studied the heat-induced denaturation and aggregation of β-lactoglobulin as a function of pH in the range 6.0-8.0; Law et al 11 studied the effect of pH on thermal denaturation of four main whey protein fractions in skim milk by gel permeation fast protein liquid chromatography; Guo et al 12 studied the enthalpy-entropy compensation relationship in protein unfolding; Antonucci et al 13 studied the minor conformational changes of a doxorubicin-peptide conjugate; Lumry 14 studied the uses of enthalpy-entropy compensation in protein molecule research; Hearn et al 15 studied the binding behavior and conformational properties of globular proteins in the presence of immobilized non-polar ligands used in reversed-phase liquid chromatography; Naletova et al 16 studied the interaction between GroEL and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) with different denatured forms.…”