No cofactor effect on equilibrium unfolding of Desulfovibrio desulfuricans flavodoxin

Apiyo, David; Guidry, Jessie; Wittung-Stafshede, Pernilla

doi:10.1016/s0167-4838(00)00032-7

Cited by 26 publications

(66 citation statements)

References 32 publications

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“…Wittung-Stafshede and co-workers (23) propose that FMN binding has no effect on the denaturant-induced equilibrium unfolding of Desulfovibrio desulfuricans flavodoxin, whereas the presence of FMN speeds up the folding of this protein (24). The resulting thermodynamic conflict is handled by WittungStafshede and co-workers by assuming that unfolded D. desulfuricans flavodoxin binds to FMN with nanomolar affinity, which is a rather remarkable assumption.…”

Section: Denaturant-induced Unfolding Shows That a Vinelandii Holoflmentioning

confidence: 99%

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Bollen¹,

Nabuurs

Berkel

et al. 2005

Journal of Biological Chemistry

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Section: Denaturant-induced Unfolding Shows That a Vinelandii Holoflmentioning

confidence: 99%

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Bollen¹,

Nabuurs

Berkel

et al. 2005

Journal of Biological Chemistry

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“…Whether or not a cofactor stabilizes the protein depends on the particular polypeptide chain. For instance, the cytochrome b562 of E. coli is stabilized by 14 kJ mol -1 when the heme cofactor is present [42,43], but the stability of the flavodoxin of Desulfovibrio desulfuricans was not affected by the presence of flavin [44]. (Table 1), which would yield a ∆ASA = 15186 Å 2 .…”

Section: Equilibrium Unfolding Of Smeldhp Involves a Non-native Monommentioning

confidence: 99%

Metal-triggered changes in the stability and secondary structure of a tetrameric dihydropyrimidinase: A biophysical characterization

Martínez‐Rodríguez

Encinar

Hurtado-Gómez

et al. 2009

Biophysical Chemistry

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT 3 ABSTRACTDihydropyrimidinase is involved in the reductive pathway of pyrimidine degradation, catalysing the reversible hydrolysis of the cyclic amide bond (-CO-NH-) of 5,6-dihydrouracil and 5,6-dihydrothymine to the corresponding N-carbamoyl-β-amino acids. This enzyme is an attractive candidate for commercial production of D-amino acids, which are used in the production of semisynthetic β-lactams, antiviral agents, artificial sweeteners, peptide hormones and pesticides. We have obtained the crystal structure of the dihydropyrimidinase from Sinorhizobium meliloti (SmelDhp) in the presence of zinc ions, but we have not been able to obtain good diffracting crystals in its absence. Then, the role of the ion in the structure of the protein, and in its stability, remains to be elucidated. In this work, the stability and the structure of SmelDhp have been studied in the absence and in the presence of zinc. In its absence, the protein acquired a tetrameric functional structure at pH ~6.0, which is stable up to pH ~9.0, as concluded from fluorescence and CD.Chemical-denaturation occurred via a monomeric intermediate with non-native structure. The addition of zinc caused: (i) an increase of the helical structure, and changes in the environment of aromatic residues; and, (ii) a higher thermal stability. However, chemical-denaturation still occurred through a monomeric intermediate. This is the first hydantoinase whose changes in the stability and in the secondary structure upon addition of zinc are described and explained, and one of the few examples where the zinc exclusively alters the secondary helical structure and the environment of some aromatic residues in the protein, leaving unchanged the quaternary structure.

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“…A single folding study of D. desulfuricans (apo)flavodoxin from ATTC strain 27774 has been reported, which suggests that during its GuHCl-dependent equilibrium folding an intermediate populates. This species has a more solvent exposed tryptophan than native protein and judged by far-UV CD ellipticity at 220 nm it has native-like secondary structure (104). Folding of apoflavodoxin from D. desulfuricans strain ATTC 29577 (75 % sequence identity with ATTC 27774 protein) has been studied more extensively.…”

Section: Folding Of Other Flavodoxinsmentioning

confidence: 99%

“…For wild-type flavodoxin (ATTC strains 27774 and 29577) it was suggested that FMN binding has no effect on its GuHCl-dependent equilibrium folding (104,105). However, FMN speeds up the folding of this protein (105).…”

Section: Folding Of Other Flavodoxinsmentioning

confidence: 99%

“…However, FMN speeds up the folding of this protein (105). The researchers involved handled the resulting thermodynamic conflict by assuming that unfolded wild-type protein binds FMN with nanomolar affinity (104,105), which is a remarkable assumption. Indeed, with urea, FMN dissociates prior to protein unfolding and urea-unfolded apoflavodoxin does not interact with FMN (106).…”

Section: Folding Of Other Flavodoxinsmentioning

confidence: 99%

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The ribosome regulates flavodoxin folding

Houwman¹

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No cofactor effect on equilibrium unfolding of Desulfovibrio desulfuricans flavodoxin

Cited by 26 publications

References 32 publications

Last In, First Out

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Metal-triggered changes in the stability and secondary structure of a tetrameric dihydropyrimidinase: A biophysical characterization

The ribosome regulates flavodoxin folding

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