Thermodynamics of Inhibitor Binding to the Catalytic Site of Glucoamylase from Aspergillus niger Determined by Displacement Titration Calorimetry

Sigurskjold, Bent W.; Berland, Carolyn R.; Svensson, Birte

doi:10.1021/bi00199a048

Cited by 71 publications

(61 citation statements)

References 52 publications

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“…Previous studies have investigated the binding affinity of acyl-CoAs for ACBP using epr spectroscopy (17) and titration microcalorimetry (22,(35)(36)(37) ranging from 4 ϫ 10 Ϫ10 to 4.5 ϫ 10 Ϫ14 M depending on acyl chain length and the method used. Measurements made by direct titration microcalorimetry range from 2.4 ϫ 10 Ϫ7 and 1.7 ϫ 10 Ϫ8 M for octanoyl-CoA and dodecanoyl-CoA with bovine ACBP (35, 36) to 8.5 ϫ 10 Ϫ11 M for palmitoyl-CoA with yeast ACBP (22).…”

Section: Resultsmentioning

confidence: 99%

Cloning, Expression, and Characterization of the acyl-CoA-binding Protein in African Trypanosomes

Milne

Ferguson

2000

Journal of Biological Chemistry

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African trypanosomes are shielded from their hosts' defenses by a coat of variant surface glycoprotein molecules, each of which is attached to the plasma membrane by a glycosylphosphatidylinositol anchor. During the later stages of glycosylphosphatidylinositol biosynthesis, myristic acid is incorporated into the anchor from the donor myristoyl-CoA by a series of unique fatty acid remodeling and exchange reactions. We have cloned and expressed a recombinant trypanosome acylCoA-binding protein that has a preference for binding relatively short chain acyl-CoAs and that has a high affinity for binding myristoyl-CoA (K d ‫؍‬ 3.5 ؋ 10This protein enhances fatty acid remodeling of glycosylphosphatidylinositol precursors in the trypanosome cell-free system. We speculate that the trypanosome acyl-CoA-binding protein plays an active role in supplying myristoyl-CoA to the fatty acid remodeling machinery in the parasite.

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Section: Resultsmentioning

confidence: 99%

Cloning, Expression, and Characterization of the acyl-CoA-binding Protein in African Trypanosomes

Milne

Ferguson

2000

Journal of Biological Chemistry

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“…The peaks of the thermograms obtained in this manner were integrated using the ORIGIN software supplied with the instrument. A nonlinear regression fitting to the isotherm was done using the CALREG (version 3.0) program (15). The fitting procedure yields the binding constant of the ligand K a , the heat of binding H, and the concentration of the binding sites (stoichiometry) N.…”

Section: Methodsmentioning

confidence: 99%

Fatty Acyl-CoA Binding Domain of the Transcription Factor FadR

DiRusso

Tsvetnitsky²,

Højrup

et al. 1998

Journal of Biological Chemistry

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“…The reason for using indirect microcalorimetry for measurement of palmitoyl-CoA binding to bovine ACBP was that the binding affinity was too high to be determined by direct microcalorimetry. The K d value for palmitoyl-CoA binding obtained by indirect titration of the octanoyl-CoA-ACBP complex with palmitoyl-CoA was calculated using the method of Sigurskjold et al [141]. However, calculation of the dissociation constant from the same data using a different method published by Hu and Eftink [142] yields a dissociation constant of 2 nM.…”

Section: Role Of Acbp In Acyl-coa Metabolism and Acyl-coa-mediated Cementioning

confidence: 99%

Role of long-chain fatty acyl-CoA esters in the regulation of metabolism and in cell signalling

Færgeman¹,

Knudsen²

1997

Biochemical Journal

629

477

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The intracellular concentration of free unbound acyl-CoA esters is tightly controlled by feedback inhibition of the acyl-CoA synthetase and is buffered by specific acyl-CoA binding proteins. Excessive increases in the concentration are expected to be prevented by conversion into acylcarnitines or by hydrolysis by acyl-CoA hydrolases. Under normal physiological conditions the free cytosolic concentration of acyl-CoA esters will be in the low nanomolar range, and it is unlikely to exceed 200 nM under the most extreme conditions. The fact that acetyl-CoA carboxylase is active during fatty acid synthesis (Ki for acyl-CoA is 5 nM) indicates strongly that the free cytosolic acyl-CoA concentration is below 5 nM under these conditions. Only a limited number of the reported experiments on the effects of acyl-CoA on cellular functions and enzymes have been carried out at low physiological concentrations in the presence of the appropriate acyl-CoA-buffering binding proteins. Re-evaluation of many of the reported effects is therefore urgently required. However, the observations that the ryanodine-senstitive Ca2+-release channel is regulated by long-chain acyl-CoA esters in the presence of a molar excess of acyl-CoA binding protein and that acetyl-CoA carboxylase, the AMP kinase kinase and the Escherichia coli transcription factor FadR are affected by low nanomolar concentrations of acyl-CoA indicate that long-chain acyl-CoA esters can act as regulatory molecules in vivo. This view is further supported by the observation that fatty acids do not repress expression of acetyl-CoA carboxylase or Δ9-desaturase in yeast deficient in acyl-CoA synthetase.

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Thermodynamics of Inhibitor Binding to the Catalytic Site of Glucoamylase from Aspergillus niger Determined by Displacement Titration Calorimetry

Cited by 71 publications

References 52 publications

Cloning, Expression, and Characterization of the acyl-CoA-binding Protein in African Trypanosomes

Cloning, Expression, and Characterization of the acyl-CoA-binding Protein in African Trypanosomes

Fatty Acyl-CoA Binding Domain of the Transcription Factor FadR

Role of long-chain fatty acyl-CoA esters in the regulation of metabolism and in cell signalling

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