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Thermal stability of bovine-brain myelin membrane

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Abstract

The thermal behaviour of bovine-brain myelin membrane has been studied by high-sensitivity differential scanning calorimetry, Fourier-transform infrared spectroscopy and thermal gel analysis. Spectroscopic results indicate that protein transitions take place between 60°C and 90°C, while thermal gel analysis has provided the thermal denaturation profiles of myelin proteolipid, DM-20 protein and the Wolfgram Fraction. An irreversible calorimetric transition centred at 80.3 ± 0.2°C with a specific enthalpy of 4.7 ± 0.6 J/g of total protein has been assigned to the thermal denaturation of myelin proteolipid and DM-20 protein. The effects of the myelin storage conditions, scan rate, ionic strength and pH on this calorimetric transition have also been investigated. The thermal transition of the proteolipid practically disappears after treatment of the myelin with different amounts of chloroform-methanol 2:1 (v/v), a treatment which is generally used in proteolipid purification. On the other hand, the addition of several detergents to myelin only causes minor modifications to this transition, which then occurs at about 70°C, with a specific enthalpy of between 2.5 and 3.6 J/g of total protein. These results appear to show that detergents preserve the native conformation of the proteolipid far more than do organic solvents. Hence the use of detergents would seem to be the appropriate method for proteolipid purification.

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Abbreviations

DSC:

Differential scanning calorimetry

TGA:

Thermal gel analysis

FTIR:

Fourier-transform infrared spectroscopy

PLP:

Proteolipid protein

MBP:

Myelin basic protein

DM-20:

Protein DM-20

WF:

Wolfgram fraction

BSA:

Bovine serum albumine

SDS:

Sodium dodecyl sulfate

ANSA:

4-amino-3-hydroxynaphthalene-1-sulphonic acid

OG:

β-d-glucopyranoside

PAGE:

Polyacrylamide gel electrophoresis

Chaps:

3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate

CNS:

Central nervous system

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Authors and Affiliations

  1. Departamento de Química Fisica (Facultad de Ciencias) e Instituto de Biotecnología, Universidad de Granada, E-18071, Granada, Spain

    J. Ruiz-Sanz, O. Lopez-Mayorga & P. L. Mateo

  2. Departamento de Química Fisica II, Facultad de Farmacia, Universidad Complutense de Madrid, E-28039, Madrid, Spain

    J. Ruiz-Cabello & M. Cortijo

Authors
  1. J. Ruiz-Sanz
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  2. J. Ruiz-Cabello
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  3. O. Lopez-Mayorga
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  4. M. Cortijo
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  5. P. L. Mateo
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Correspondence to: P. L. Mateo

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Ruiz-Sanz, J., Ruiz-Cabello, J., Lopez-Mayorga, O. et al. Thermal stability of bovine-brain myelin membrane. Eur Biophys J 21, 169–178 (1992). https://doi.org/10.1007/BF00196760

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  • DOI: https://doi.org/10.1007/BF00196760

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