Isolation and characterization of muscle membranes using surface-specific labels
- PMID: 138140
- PMCID: PMC393191
- DOI: 10.1073/pnas.74.1.34
Isolation and characterization of muscle membranes using surface-specific labels
Abstract
Membranes were purified from rat muscle by a differential centrifugation method that avoids the use of salt extraction or incubations at elevated temperature. Three populations of membrane-limited vesicles were defined having average densities of 1.112 (fraction I), 1.141 (fraction II), and 1.158 (fraction III) g/ml in a continuous sucrose gradient. Lactoperoxidase-catalyzed iodination of intact muscle prior to isolation of membranes resulted in highest specific activity in fraction I, although all fractions could be equally labeled after isolation. 125I-Labeled wheat germ agglutinin incubated at low concentration with intact muscle preferentially labeled fraction I. Parallel studies on previously isolated fractions indicated that fraction I also contained the highest concentration of potential receptors for wheat germ agglutinin. In experiments on whole muscle, concanavalin A bound predominantly to sarcolemma with slight variable binding to T-tubular and nuclear membrane but no binding to sarcoplasmic reticulum or mitochondria. Parallel binding studies with isolated membrane fragments indicated heavy binding of concanavalin A by membranes in fraction I with scattered binding in fractions II and III. Na+K+Mg2+ ATPase was specifically enriched in fraction I but was also present in fraction II in a proportion similar to 125I labeling. Ca2+ ATPase was most active in fraction II but present in significant levels in fraction I. It is concluded from these and other data that fraction I contains predominantly sarcolemma membrane, while T-tubular membrane may represent a significant component of fraction II. Ca2+ ATPase activity in fraction I is intrinsic to the sarcolemma.
Similar articles
-
Isolation and partial characterization of an acetylcholine receptor-enriched membrane fraction from skeletal muscle.J Recept Res. 1981-1982;2(5-6):503-21. doi: 10.3109/107998981809038882. J Recept Res. 1981. PMID: 6296380
-
In vitro analysis of the general properties and junctional receptor characteristics of skeletal muscle membranes. Isolation, purification, and partial characterization of sarcolemmal fragments.Proc Natl Acad Sci U S A. 1974 Jun;71(6):2435-9. doi: 10.1073/pnas.71.6.2435. Proc Natl Acad Sci U S A. 1974. PMID: 4276296 Free PMC article.
-
Isolation and characterization of distinct domains of sarcolemma and T-tubules from rat skeletal muscle.Biochem J. 1995 Apr 1;307 ( Pt 1)(Pt 1):273-80. doi: 10.1042/bj3070273. Biochem J. 1995. PMID: 7536412 Free PMC article.
-
One-step isolation and characterization of nuclear membranes.Philos Trans R Soc Lond B Biol Sci. 1974 Jul 25;268(891):101-8. doi: 10.1098/rstb.1974.0018. Philos Trans R Soc Lond B Biol Sci. 1974. PMID: 4155085 Review. No abstract available.
-
[Enzymatic properties in muscle membranes].Ukr Biokhim Zh. 1975 Sep-Oct;47(5):619-34. Ukr Biokhim Zh. 1975. PMID: 128174 Review. Russian.
Cited by
-
Biochemical properties of isolated transverse tubular membranes.J Bioenerg Biomembr. 1989 Apr;21(2):163-213. doi: 10.1007/BF00812068. J Bioenerg Biomembr. 1989. PMID: 2473982 Review.
-
Characteristics of saxitoxin binding to the sodium channel of sarcolemma isolated from rat skeletal muscle.J Physiol. 1979 Oct;295:383-96. doi: 10.1113/jphysiol.1979.sp012975. J Physiol. 1979. PMID: 42783 Free PMC article.
-
Microsomal T system: a stereological analysis of purified microsomes derived from normal and dystrophic skeletal muscle.J Cell Biol. 1979 Oct;83(1):33-46. doi: 10.1083/jcb.83.1.33. J Cell Biol. 1979. PMID: 511940 Free PMC article.
-
Incorporation of amino acids into soluble and membrane protein fractions of dystrophic hamsters.Biochem J. 1980 Aug 15;190(2):341-8. doi: 10.1042/bj1900341. Biochem J. 1980. PMID: 7470053 Free PMC article.
-
Tri-partite complex for axonal transport drug delivery achieves pharmacological effect.BMC Neurosci. 2010 Jan 20;11:8. doi: 10.1186/1471-2202-11-8. BMC Neurosci. 2010. PMID: 20085661 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Miscellaneous