Alterations in the natural abundance /sup 13/C NMR spectra of skeletal muscle membranes depending on the extraction medium of muscle
Minced rat muscles were extracted with NaCl solution with additions: 1) 10 mM EDTA, or 2) 2 mM MgCl/sub 2/, or 3) None (final ionic strength equivalent to 150 mM NaCl, pH 7.0). The total muscle membranes were pelleted between 1500 and 252,000 x g. All membranes were washed several times by homogenization with 0.05 M NaCl, pH 6.1, and centrifugation at 252,000 x g. The final pellets were suspended in /sup 2/H/sub 2/O to contain 150 mg membrane protein/ml in 30% /sup 2/H/sub 2/O. /sup 13/C NMR spectra were recorded at 50.3 MHz, under fully relaxed conditions. The spectral pattern varied according to the extraction medium of muscle: Relative to None addition, EDTA caused a decrease in the intensity of the polyunsaturated carbon resonance (128.6 ppm), whereas with MgCl/sub 2/ the opposite effect was found. Metal analysis, after combustion of the membranes, showed 1.1-1.6 mol Ca/sup 2 +/ per 10/sup 5/ g protein in None and Mgcl/sub 2/ membranes, and 1000-times less Ca/sup 2 +/ in EDTA membranes. The Mg/sup 2 +/ content of the membranes was not affected by EDTA, however, it was increased 5-fold when MgCl/sub 2/ was present in the extraction medium. Thus, the results indicate that membrane-bound Ca/sup 2 +/ and Mg/sup 2 +/ alter the conformation of membrane-phospholipis.
- Research Organization:
- Univ. of Illinois, Chicago
- OSTI ID:
- 5496572
- Report Number(s):
- CONF-8604222-; TRN: 86-026687
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CELL MEMBRANES
NMR SPECTRA
CALCIUM COMPOUNDS
CARBON 13
EDTA
MAGNESIUM CHLORIDES
MUSCLES
PHOSPHOLIPIDS
SOLVENT EXTRACTION
ALKALINE EARTH METAL COMPOUNDS
AMINO ACIDS
CARBON ISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHELATING AGENTS
CHLORIDES
CHLORINE COMPOUNDS
ESTERS
EVEN-ODD NUCLEI
EXTRACTION
HALIDES
HALOGEN COMPOUNDS
ISOTOPES
LIGHT NUCLEI
LIPIDS
MAGNESIUM COMPOUNDS
MEMBRANES
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
SEPARATION PROCESSES
SPECTRA
STABLE ISOTOPES
550601* - Medicine- Unsealed Radionuclides in Diagnostics