Subfractionation of heart tissue: The "in sequence" myofilament protein extraction of myocardial tissue

Lesley A. Kane, Irina Neverova, Jennifer E. Van Eyk

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Proteomic analysis of heart tissue is complicated by the large dynamic range of its proteins. The most abundant proteins are the myofilament proteins, which comprise the contractile apparatus. This chapter describes a protocol for fractionation of heart tissue that extracts the myofilament proteins into a separate sample fraction, allowing analysis of lower-abundance proteins. Importantly, this is performed in a manner that is compatible with two-dimensional electrophoresis and high-performance liquid chromatography, two of main technologies of proteomics. The method produces three fractions based on solubility at different pHs: (1) cytoplasmic-enriched extract (neutral pH), (2) myofilament-enriched extract (acidic pH), and (3) membrane protein-enriched pellet. Fractionation of heart tissue in this manner provides the basis for in-depth proteomic analysis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages87-90
Number of pages4
Volume357
DOIs
StatePublished - Feb 5 2007
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume357
ISSN (Print)10643745

Fingerprint

Myofibrils
Proteomics
Proteins
Tissue Extracts
Solubility
Electrophoresis
Membrane Proteins
High Pressure Liquid Chromatography
Technology

Keywords

  • Myofilaments
  • Striated muscle
  • Subproteome fractionation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Kane, L. A., Neverova, I., & Van Eyk, J. E. (2007). Subfractionation of heart tissue: The "in sequence" myofilament protein extraction of myocardial tissue. In Methods in Molecular Biology (Vol. 357, pp. 87-90). (Methods in Molecular Biology; Vol. 357). https://doi.org/10.1385/1-59745-214-9:87

Subfractionation of heart tissue : The "in sequence" myofilament protein extraction of myocardial tissue. / Kane, Lesley A.; Neverova, Irina; Van Eyk, Jennifer E.

Methods in Molecular Biology. Vol. 357 2007. p. 87-90 (Methods in Molecular Biology; Vol. 357).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kane, LA, Neverova, I & Van Eyk, JE 2007, Subfractionation of heart tissue: The "in sequence" myofilament protein extraction of myocardial tissue. in Methods in Molecular Biology. vol. 357, Methods in Molecular Biology, vol. 357, pp. 87-90. https://doi.org/10.1385/1-59745-214-9:87
Kane LA, Neverova I, Van Eyk JE. Subfractionation of heart tissue: The "in sequence" myofilament protein extraction of myocardial tissue. In Methods in Molecular Biology. Vol. 357. 2007. p. 87-90. (Methods in Molecular Biology). https://doi.org/10.1385/1-59745-214-9:87
Kane, Lesley A. ; Neverova, Irina ; Van Eyk, Jennifer E. / Subfractionation of heart tissue : The "in sequence" myofilament protein extraction of myocardial tissue. Methods in Molecular Biology. Vol. 357 2007. pp. 87-90 (Methods in Molecular Biology).
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