Removal of bound Triton X-100 from purified bovine heart cytochrome bc1

Rastislav Varhač; Neal C. Robinson; Andrej Musatov

doi:10.1016/j.ab.2009.08.042

Removal of bound Triton X-100 from purified bovine heart cytochrome bc₁

Rastislav Varhač, Neal C. Robinson, Andrej Musatov

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

Cytochrome bc₁ isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc₁ is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc₁ was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc₁ retained nearly full activity, absorption spectra, subunit, and phospholipid composition.

Original language	English (US)
Pages (from-to)	268-270
Number of pages	3
Journal	Analytical Biochemistry
Volume	395
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2009.08.042
State	Published - Dec 15 2009
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Removal of bound Triton X-100 from purified bovine heart cytochrome bc1",

abstract = "Cytochrome bc1 isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc1 is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc1 retained nearly full activity, absorption spectra, subunit, and phospholipid composition.",

author = "Rastislav Varha{\v c} and Robinson, {Neal C.} and Andrej Musatov",

note = "Funding Information: This work was supported by grants from UTHSCSA (URC Grant 128240 to A.M.), the National Institutes of Health (NIH GM024795 to N.C.R.), and the Robert A. Welch Foundation ( AQ1481 to N.C.R.). The authors thank LeAnn K. Robinson for editorial help in preparing the manuscript.",

year = "2009",

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doi = "10.1016/j.ab.2009.08.042",

language = "English (US)",

volume = "395",

pages = "268--270",

journal = "Analytical Biochemistry",

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T1 - Removal of bound Triton X-100 from purified bovine heart cytochrome bc1

AU - Varhač, Rastislav

AU - Robinson, Neal C.

AU - Musatov, Andrej

N1 - Funding Information: This work was supported by grants from UTHSCSA (URC Grant 128240 to A.M.), the National Institutes of Health (NIH GM024795 to N.C.R.), and the Robert A. Welch Foundation ( AQ1481 to N.C.R.). The authors thank LeAnn K. Robinson for editorial help in preparing the manuscript.

PY - 2009/12/15

Y1 - 2009/12/15

N2 - Cytochrome bc1 isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc1 is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc1 retained nearly full activity, absorption spectra, subunit, and phospholipid composition.

AB - Cytochrome bc1 isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc1 is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc1 was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc1 retained nearly full activity, absorption spectra, subunit, and phospholipid composition.

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

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ER -

Removal of bound Triton X-100 from purified bovine heart cytochrome bc₁

Abstract

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