Fourier transform infrared spectroscopic studies on gastric H+/K+-ATPase

Mitchell, Robert E.; Haris, Parvez I.; Fallowfield, Colin; Keeling, David J.; Chapman, D.

doi:10.1016/0005-2736(88)90210-6

Cited by 33 publications

(15 citation statements)

References 35 publications

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“…Gresalfi and Wallace [17] suggested a structural change affecting 7% of the Na + ,K + ‐ATPase on the basis of CD measurements. Later, Chetverin and Brazhnikov [18], using infrared spectroscopy, came to the conclusion that the change affected less than 2% of the residues, in agreement with a similar estimation obtained for the H + ,K + ‐ATPase [19].…”

supporting

confidence: 76%

Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Raussens

Pézolet

Ruysschaert

et al. 1999

European Journal of Biochemistry

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+ were investigated by different spectroscopic approaches. No significant secondary-structure change or secondary-structure reorientation with respect to the membrane plane could be measured by attenuated total reflection Fourier transform infrared spectroscopy of oriented films. Circular dichroism and Raman spectra obtained on tubulovesicle suspensions indicated no significant secondary structure or tyrosine and tryptophan side-chain environment changes in tubulovesicle suspensions. The smallest observable structural changes are discussed in term of the number of amino-acid residues involved for each technique.

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supporting

confidence: 76%

Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Raussens

Pézolet

Ruysschaert

et al. 1999

European Journal of Biochemistry

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“…By far the most widely used of these bands is the amide I band (2)(3)(4)(5)(10)(11)(12), closely followed by the amide Π. The amide ΠΙ (33,34) and amide A (35,36) bands have also been successfully used. The usefulness of the Amide HI band for protein secondary structural analysis has been most effectively demonstrated by Singh and co-workers (ref.…”

Section: Information On Peptide Secondary Structure From Ftir Spectramentioning

confidence: 99%

Fourier Transform Infrared Spectroscopic Studies of Peptides: Potentials and Pitfalls

Haris

1999

ACS Symposium Series

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This article presents a discussion of the advantages and drawbacks of FTIR spectroscopy for structural characterisation of peptides. FTIR spectra of small peptides can be highly complex and extremely difficult to interpret. Amide bands associated with different secondary structural elements in peptides can show large deviations from those observed with large water-soluble proteins. This is partly due to the fact small peptides are often highly flexible and are solvated to a greater extent compared to globular proteins. Peptide aggregation can also result in the appearance of additional bands in the amide I region that can be erroneously attributed to peptide structure. These are amongst a host of factors that needs to be taken into consideration when interpreting infrared spectra of peptides.Infrared spectroscopy is one of the earliest techniques to be used for protein conformational analysis (1). From the frequency, intensity and width of the amide bands in the infrared spectrum of a protein, it is possible to obtain highly detailed information on its structure. A survey of the current literature clearly indicates the popularity of Fourier transform infrared (FTIR) spectroscopy as a tool for structural characterisation of peptides and proteins (for reviews see ref [2][3][4][5][6][7][8][9][10][11][12]. This popularity can be attributed to a number of factors, including the ability to use this technique for recording spectra of biomolecules in diverse environments. It enables conformational analysis of a peptide to be carried out in aqueous solution, crystals, organic solvents, detergents, micelles as well as in phospholipid membranes. As such FTIR spectroscopy is one of the few techniques that can be used to monitor the role of the surrounding environment on protein conformation. This is important in light of increasing evidence 54

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“…The changes in secondary or tertiary structure of H 1 /K 1 -ATPase occurring during its catalytic cycle are not known. According to Mitchell and coworkers [9], the E1±E2 transition does not involve significant changes of the secondary structure content, i.e. the distribution of a helix or b sheet stays unmodified.…”

mentioning

confidence: 98%

Monitoring of secondary and tertiary structure changes in the gastric H⁺/K⁺‐ATPase by infrared spectroscopy

Scheirlinckx

Buchet

Ruysschaert

et al. 2001

European Journal of Biochemistry

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Conformational changes occurring in the catalytic cycle of the H 1 /K 1 -ATPase were monitored by Fourier transform infrared spectroscopy (FTIR). Caged compounds were used to release ATP, in the presence of Ca 21 , to induce the transition between the E1 and E1-P conformation of the H 1 /K 1 -ATPase. In addition to bands associated with the photolysis of the caged compounds, some peaks of the difference infrared spectra were associated with changes in secondary structure and modifications of the ionization in the side chains of amino-acid residues (Glu or Asp). These changes were specific to the reaction between the ligand and the enzyme. We estimated that 39 amino acids changed their secondary structure during the reaction and four amino-acid residues were deprotonated. Similar spectral changes appeared when ADP was released from its precursor. The release of P i from the same caged molecule did not induce similar changes. Changes in tertiary structure occurring during the binding of adenosine and phosphorylation of the enzyme were demonstrated by recording hydrogen/deuterium exchange kinetics by attenuated total reflectance FTIR spectroscopy (ATR-FTIR). At least 129 amide protons were involved in a tertiary structure change induced by ATP. This suggested that secondary structure change transduced a much larger tertiary structure modification.Keywords: H 1 /K 1 -ATPase; caged-compounds; catalytic cycle; ATR-FTIR spectroscopy.The gastric H 1 /K 1 -ATPase is the electroneutral ion pump responsible for acid secretion in the gastric mucosa. In this process, cytosolic H 1 is exchanged for extracellular K 1 . The gastric H 1 /K 1 -ATPase is a member of the P-ATPase family and shares many features, including sequence homologies and enzymatic mechanisms with other members such as the Ca 21 -ATPase [1], and especially the Na 1 /K 1 -ATPase [2]. Enzymes of this large family are characterized by the formation of a covalent aspartylphosphate group during their catalytic cycle and the transition between two major conformations called E1 and E2.From a structural point of view, this enzyme is composed of a catalytic a subunit (114 kDa) and a glycosylated b subunit (33 kDa). The a subunit, a membrane multispanning polypeptide, is responsible for the coupling of ATP hydrolysis with ion transport across the membrane. The b subunit, a single membrane spanning peptide, is required for the proper assembly and targeting of the entire protein. The a subunit of the H 1 /K 1 -ATPase consists of 1034 amino-acid residues [2]. It contains the phosphorylation site (Asp385) and Lys517, which is preferentially labeled by fluorescein 5 H -isothiocyanate (FITC). The models based on hydrophobicity profiles and different experimental results predict 10 membrane spanning segments for the a subunit. Although segments M9 and M10 have never been isolated after proteolysis, genetics experiments support this view [2,3]; it is also consistent with the recently published structure of the Ca 21 -ATPase [4]. The b subunit consists of 290 amino-acid residu...

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Fourier transform infrared spectroscopic studies on gastric H+/K+-ATPase

Cited by 33 publications

References 35 publications

Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Fourier Transform Infrared Spectroscopic Studies of Peptides: Potentials and Pitfalls

Monitoring of secondary and tertiary structure changes in the gastric H⁺/K⁺‐ATPase by infrared spectroscopy

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Fourier transform infrared spectroscopic studies on gastric H+/K+-ATPase

Cited by 33 publications

References 35 publications

Structural difference in the H+,K+‐ATPase between the E1 and E2 conformations

Structural difference in the H+,K+‐ATPase between the E1 and E2 conformations

Fourier Transform Infrared Spectroscopic Studies of Peptides: Potentials and Pitfalls

Monitoring of secondary and tertiary structure changes in the gastric H+/K+‐ATPase by infrared spectroscopy

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Product

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Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Structural difference in the H⁺,K⁺‐ATPase between the E1 and E2 conformations

Monitoring of secondary and tertiary structure changes in the gastric H⁺/K⁺‐ATPase by infrared spectroscopy