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Evidence from Raman Spectroscopy That InhA, the Mycobacterial Enoyl Reductase, Modulates the Conformation of the NADH Cofactor to Promote Catalysis.

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Title: Evidence from Raman Spectroscopy That InhA, the Mycobacterial Enoyl Reductase, Modulates the Conformation of the NADH Cofactor to Promote Catalysis.
Authors: Bell, Alasdair F.1, Stratton, Christopher F.2, Xujie Zhang1, Novichenok, Polina1, Jaye, Andrew A.1, Nair, Pravin A.3, Parikh, Sapan1, Rawat, Richa1, Tonge, Peter J.1 peter.tonge@sunysb.edu
Source: Journal of the American Chemical Society. 5/23/2007, Vol. 129 Issue 20, p6425-6431. 7p.
Subjects: Raman spectroscopy, Size reduction of materials, Raman effect, Nanoparticles, Physical & theoretical chemistry, Chemistry
Abstract: InhA, the enoyl reductase from Mycobacterium tuberculosis, catalyzes the NADH-dependent reduction of trans-2-enoyl-ACPs. In the present work, Raman spectroscopy has been used to identify catalytically relevant changes in the conformation of the nicotinamide ring that occur when NADH binds to InhA. For 4(S)-NADD, there is an 11 cm-1 decrease in the wavenumber of the C4-D stretching band (νC-D) and a 50% decrease in the width of this band upon binding to InhA. While a similar reduction in line width is observed for the corresponding band arising from 4(R)-NADD, νC-D for this isomer increases 34 cm-1 upon binding to InhA. These changes in νC-D indicate that the nicotinamide ring adopts a bound conformation in which the 4(S)C-D bond is in a pseudoaxial orientation. Mutagenesis of F149, a conserved active site residue close to the cofactor, demonstrates that this enzyme-induced modulation in cofactor structure is directly linked to catalysis. In contrast to the wild-type enzyme, Raman spectra of NADD bound to F149A InhA resemble those of NADD in solution. Consequently, F149A is no longer able to optimally position the cofactor for hydride transfer, which correlates with the 30-fold decrease in kcat and 2-fold increase in D(V/KNADH) caused by this mutation. These studies thus substantiate the proposal that hydride transfer is promoted by pseudoaxial positioning of the NADH pro-4S bond, and indicate that catalysis of substrate reduction by InhA results, in part, from correct orientation of the cofactor in the ground state. [ABSTRACT FROM AUTHOR]
Copyright of Journal of the American Chemical Society is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Evidence from Raman Spectroscopy That InhA, the Mycobacterial Enoyl Reductase, Modulates the Conformation of the NADH Cofactor to Promote Catalysis.
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  Data: <searchLink fieldCode="AR" term="%22Bell%2C+Alasdair+F%2E%22">Bell, Alasdair F.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Stratton%2C+Christopher+F%2E%22">Stratton, Christopher F.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Xujie+Zhang%22">Xujie Zhang</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Novichenok%2C+Polina%22">Novichenok, Polina</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Jaye%2C+Andrew+A%2E%22">Jaye, Andrew A.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Nair%2C+Pravin+A%2E%22">Nair, Pravin A.</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Parikh%2C+Sapan%22">Parikh, Sapan</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Rawat%2C+Richa%22">Rawat, Richa</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Tonge%2C+Peter+J%2E%22">Tonge, Peter J.</searchLink><relatesTo>1</relatesTo><i> peter.tonge@sunysb.edu</i>
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  Data: <searchLink fieldCode="DE" term="%22Raman+spectroscopy%22">Raman spectroscopy</searchLink><br /><searchLink fieldCode="DE" term="%22Size+reduction+of+materials%22">Size reduction of materials</searchLink><br /><searchLink fieldCode="DE" term="%22Raman+effect%22">Raman effect</searchLink><br /><searchLink fieldCode="DE" term="%22Nanoparticles%22">Nanoparticles</searchLink><br /><searchLink fieldCode="DE" term="%22Physical+%26+theoretical+chemistry%22">Physical & theoretical chemistry</searchLink><br /><searchLink fieldCode="DE" term="%22Chemistry%22">Chemistry</searchLink>
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  Data: InhA, the enoyl reductase from Mycobacterium tuberculosis, catalyzes the NADH-dependent reduction of trans-2-enoyl-ACPs. In the present work, Raman spectroscopy has been used to identify catalytically relevant changes in the conformation of the nicotinamide ring that occur when NADH binds to InhA. For 4(S)-NADD, there is an 11 cm-1 decrease in the wavenumber of the C4-D stretching band (νC-D) and a 50% decrease in the width of this band upon binding to InhA. While a similar reduction in line width is observed for the corresponding band arising from 4(R)-NADD, νC-D for this isomer increases 34 cm-1 upon binding to InhA. These changes in νC-D indicate that the nicotinamide ring adopts a bound conformation in which the 4(S)C-D bond is in a pseudoaxial orientation. Mutagenesis of F149, a conserved active site residue close to the cofactor, demonstrates that this enzyme-induced modulation in cofactor structure is directly linked to catalysis. In contrast to the wild-type enzyme, Raman spectra of NADD bound to F149A InhA resemble those of NADD in solution. Consequently, F149A is no longer able to optimally position the cofactor for hydride transfer, which correlates with the 30-fold decrease in kcat and 2-fold increase in D(V/KNADH) caused by this mutation. These studies thus substantiate the proposal that hydride transfer is promoted by pseudoaxial positioning of the NADH pro-4S bond, and indicate that catalysis of substrate reduction by InhA results, in part, from correct orientation of the cofactor in the ground state. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Journal of the American Chemical Society is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1021/ja068219m
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        Text: English
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        PageCount: 7
        StartPage: 6425
    Subjects:
      – SubjectFull: Raman spectroscopy
        Type: general
      – SubjectFull: Size reduction of materials
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      – SubjectFull: Raman effect
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      – SubjectFull: Nanoparticles
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      – SubjectFull: Physical & theoretical chemistry
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      – SubjectFull: Chemistry
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      – TitleFull: Evidence from Raman Spectroscopy That InhA, the Mycobacterial Enoyl Reductase, Modulates the Conformation of the NADH Cofactor to Promote Catalysis.
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              Text: 5/23/2007
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