DOI: 10.1083/jcb.96.4.1008. Purification of morphologically intact triad structures from skeletal muscle.

Purification of morphologically intact triad structures from skeletal muscle.

10.1083/jcb.96.4.1008

Crossref journal-article

Rockefeller University Press

The Journal of cell biology (291)

Abstract

A procedure has been devised for isolation of triads (t-tubule/sarcoplasmic reticulum (SR) junctional complexes) from rabbit skeletal muscle. The procedure consists of preparation of a heavy microsomal fraction followed by two sequential 90-min sucrose gradient centrifugations to enrich the triads. A pyrophosphate/phosphate/magnesium buffer system was introduced to decrease aggregation in order to achieve effective separation. The preparation time is 12 h. Some differences between purified triads isolated by two variants of this method are noted. The purity of the triad fractions has been estimated by particle counting to be in the vicinity of 50%. There is good retention of morphology and Ca++-loading activity and enrichment in Na+,K+-ATPase and adenylate cyclase. The triads are practically devoid of contractile elements, mitochondria, and free plasmalemma, and low in content of light SR. The method for obtaining enriched triads is reproducible, and sufficient yields are obtained for structural, biochemical, and functional characterization.

Bibliography

Mitchell, R. D., Palade, P., & Fleischer, S. (1983). Purification of morphologically intact triad structures from skeletal muscle. The Journal of Cell Biology, 96(4), 1008â1016.

Authors 3

R D Mitchell (first)
P Palade (additional)
S Fleischer (additional)

References 0 Referenced 79

None

Dates

Type	When
Created	21 years, 3 months ago (May 14, 2004, 7:04 p.m.)
Deposited	2 years, 1 month ago (July 24, 2023, 1:09 p.m.)
Indexed	1 month, 3 weeks ago (July 8, 2025, 7:30 a.m.)
Issued	42 years, 5 months ago (April 1, 1983)
Published	42 years, 5 months ago (April 1, 1983)
Published Online	42 years, 5 months ago (April 1, 1983)
Published Print	42 years, 5 months ago (April 1, 1983)

Funders 0

None

BibTeX

@article{Mitchell_1983, title={Purification of morphologically intact triad structures from skeletal muscle.}, volume={96}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.96.4.1008}, DOI={10.1083/jcb.96.4.1008}, number={4}, journal={The Journal of cell biology}, publisher={Rockefeller University Press}, author={Mitchell, R D and Palade, P and Fleischer, S}, year={1983}, month=apr, pages={1008–1016} }

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