Comparison of .alpha.-lactalbumin and lysozyme using vibrational circular dichroism. Evidence for a difference in crystal and solution structures

Urbanová, Marie; Dukor, Rina K.; Pančoška, Petr; Gupta, Vikram; Keiderling, Timothy A.

doi:10.1021/bi00107a016

Cited by 54 publications

(33 citation statements)

References 36 publications

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“…1B, broken traces). The infrared spectrum of the holoprotein in deuterated buffer displays well resolved bands at 1650 and 1640 cm Ϫ1 which have been assigned to ␣-helical segments and fully hydrated, extended chains connecting different types of secondary structure elements, respectively (20,21). Minor components appearing at 1630 and 1675 cm Ϫ1 indicate the presence of ␤-structures and turns, respectively.…”

Section: Methodsmentioning

confidence: 99%

Binding of Molten Globule-like Conformations to Lipid Bilayers

Bañuelos

Muga

1995

Journal of Biological Chemistry

114

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The effect of membrane binding on the structure and stability of several conformers of ␣-lactalbumin was studied by infrared spectroscopy, circular dichroism, and fluorescence spectroscopy. In solution, under experimental conditions where all conformers interact with negatively charged membranes, they show significant conformational differences. However, binding to negatively charged membranes, which causes considerable changes in the structure of these conformers, leads to a remarkably similar protein conformation. The membrane-associated conformations are characterized by 1) a high helical content, greater than any of those found in solution, 2) a lack of stable tertiary structure, and 3) the disappearance of their thermotropic transition. These observations indicate that association with negatively charged membranes induces a conformational change within ␣-lactalbumin to a flexible, molten globule-like state.

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Section: Methodsmentioning

confidence: 99%

Binding of Molten Globule-like Conformations to Lipid Bilayers

Bañuelos

Muga

1995

Journal of Biological Chemistry

114

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“…Recently, Urbanova et al (1991), based on vibrational circular dichroism studies of HEWL and BLA, found evidence that the secondary structure of a-lactalbumins may differ in solution and in the crystalline state. The NMR study reported here suggests that the tertiary structures as reflected in the environments of the aromatic residues in a-lactalbumins in solution and in the crystalline state are highly similar.…”

Section: Comparisons With Previous Workmentioning

confidence: 99%

¹H‐NMR assignments and local environments of aromatic residues in bovine, human and guinea pig variants of α‐lactalbumin

Alexandrescu

Broadhurst

Wormald

et al. 1992

European Journal of Biochemistry

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'H-NMR assignments have been defined for the aromatic-ring protons of the bovine, guinea pig and human variants of a-lactalbumin. Spin-system networks were identified by means of doublequantum-filtered two-dimensional J-correlated spectroscopy and two-dimensional relayed coherence spectroscopy data. Analysis of two-dimensional nuclear-Overhauser-enhancement spectroscopy data of the proteins indicated that in each case two clusters of aromatic residues exist. The two clusters are also evident in the crystal structure of the human protein, and this evidence, in conjunction with sequence differences between the three proteins, permitted sequence-specific assignments to be made for the majority of aromatic residues. Remaining ambiguities in the assignments could be resolved by analysis of photochemically induced dynamic nuclear polarization (PCIDNP) effects. Comparison of the PCIDNP spectra of the three proteins indicated the presence of only minor differences in the surface exposure of conserved aromatic residues. Taken together, these results indicate that the environments of the conserved aromatic residues in bovine, guinea pig and human a-lactalbumin in solution are very similar to each other, and that the solution and the crystal forms of at least the human protein are similar.

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“…Also, numerous lattice contacts at the protein-protein interfaces can subtly alter the crystal structure from the solution structure. Examples are alpha-lactalbumin [29], myoglobin [30] and aspartate transcarbamylase [31]. Finally, it is known that crystallization may not demonstrate structural variability as in the case of intestinal fatty-acid-binding apoprotein where X-ray crystallization did not demonstrate the variability of backbone structure [32].…”

Section: Discussionmentioning

confidence: 99%

Stereoselectivity of Isoflurane in Adhesion Molecule Leukocyte Function-Associated Antigen-1

Pereira

Eckenhoff

et al. 2014

PLoS ONE

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BackgroundIsoflurane in clinical use is a racemate of S- and R-isoflurane. Previous studies have demonstrated that the effects of S-isoflurane on relevant anesthetic targets might be modestly stronger (less than 2-fold) than R-isoflurane. The X-ray crystallographic structure of the immunological target, leukocyte function-associated antigen-1 (LFA-1) with racemic isoflurane suggested that only S-isoflurane bound specifically to this protein. If so, the use of specific isoflurane enantiomers may have advantage in the surgical settings where a wide range of inflammatory responses is expected to occur. Here, we have further tested the hypothesis that isoflurane enantioselectivity is apparent in solution binding and functional studies.MethodsFirst, binding of isoflurane enantiomers to LFA-1 was studied using 1-aminoanthracene (1-AMA) displacement assays. The binding site of each enantiomer on LFA-1 was studied using the docking program GLIDE. Functional studies employed the flow-cytometry based ICAM binding assay.ResultsBoth enantiomers decreased 1-AMA fluorescence signal (at 520 nm), indicating that both competed with 1-AMA and bound to the αL I domain. The docking simulation demonstrated that both enantiomers bound to the LFA-1 “lovastatin site.” ICAM binding assays showed that S-isoflurane inhibited more potently than R-isoflurane, consistent with the result of 1-AMA competition assay.ConclusionsIn contrast with the x-ray crystallography, both enantiomers bound to and inhibited LFA-1. S-isoflurane showed slight preference over R-isoflurane.

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Comparison of .alpha.-lactalbumin and lysozyme using vibrational circular dichroism. Evidence for a difference in crystal and solution structures

Cited by 54 publications

References 36 publications

Binding of Molten Globule-like Conformations to Lipid Bilayers

Binding of Molten Globule-like Conformations to Lipid Bilayers

¹H‐NMR assignments and local environments of aromatic residues in bovine, human and guinea pig variants of α‐lactalbumin

Stereoselectivity of Isoflurane in Adhesion Molecule Leukocyte Function-Associated Antigen-1

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Comparison of .alpha.-lactalbumin and lysozyme using vibrational circular dichroism. Evidence for a difference in crystal and solution structures

Cited by 54 publications

References 36 publications

Binding of Molten Globule-like Conformations to Lipid Bilayers

Binding of Molten Globule-like Conformations to Lipid Bilayers

1H‐NMR assignments and local environments of aromatic residues in bovine, human and guinea pig variants of α‐lactalbumin

Stereoselectivity of Isoflurane in Adhesion Molecule Leukocyte Function-Associated Antigen-1

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¹H‐NMR assignments and local environments of aromatic residues in bovine, human and guinea pig variants of α‐lactalbumin