Hydrophobic interaction chromatography of myosin fragments: Potential use in purification

Julian Borejdo, Shoshana Linder, Moshe M. Werber

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Myosin fragments were fractionated on columns of the hydrophobic gel phenyl-Sepharose CL-4B. In the presence of high NaCl concentrations the fragments bound tightly to the columns; they could be eluted by decreasing the ionic strength, by increasing the pH, or by applying various concentrations of ethylene glycol. In myosin subfragment-1 (S-1), the light chains underwent partial dissociation from the heavy chain and bound separately to the column matrix. The order of strength of binding of the various species to the column was heavy chain > A1 light chain > A2 light chain > native S-1 > denatured heavy chain or S-1. Thus the hydrophobic gel appears to be able to differentiate between enzymatically active and inactive S-1. Under appropriate elution conditions it was possible to obtain S-1 preparations depleted from nicked heavy chains and with specific ATPase activities 34-130% higher than those of untreated S-1. When S-1(A2) was fractionated on phenyl-Sepharose a fivefold enrichment of the heavy chain with respect to the light chains was obtained, while the ATPase activity was equal or larger than that of the original S-1, implying that the light chains are not essential for ATPase activity. Thus, it seems that chromatography of S-1 on phenyl-Sepharose is a potentially useful method for obtaining a purified myosin heavy-chain fragment with a high ATPase specific activity.

Original languageEnglish
Pages (from-to)193-201
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume231
Issue number1
DOIs
StatePublished - 15 May 1984

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Myosins
Chromatography
Hydrophobic and Hydrophilic Interactions
Purification
Adenosine Triphosphatases
Light
varespladib methyl
Gels
Myosin Subfragments
Ethylene Glycol
Myosin Heavy Chains
Distillation columns
Ionic strength
Osmolar Concentration
phenyl-sepharose

Cite this

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title = "Hydrophobic interaction chromatography of myosin fragments: Potential use in purification",
abstract = "Myosin fragments were fractionated on columns of the hydrophobic gel phenyl-Sepharose CL-4B. In the presence of high NaCl concentrations the fragments bound tightly to the columns; they could be eluted by decreasing the ionic strength, by increasing the pH, or by applying various concentrations of ethylene glycol. In myosin subfragment-1 (S-1), the light chains underwent partial dissociation from the heavy chain and bound separately to the column matrix. The order of strength of binding of the various species to the column was heavy chain > A1 light chain > A2 light chain > native S-1 > denatured heavy chain or S-1. Thus the hydrophobic gel appears to be able to differentiate between enzymatically active and inactive S-1. Under appropriate elution conditions it was possible to obtain S-1 preparations depleted from nicked heavy chains and with specific ATPase activities 34-130{\%} higher than those of untreated S-1. When S-1(A2) was fractionated on phenyl-Sepharose a fivefold enrichment of the heavy chain with respect to the light chains was obtained, while the ATPase activity was equal or larger than that of the original S-1, implying that the light chains are not essential for ATPase activity. Thus, it seems that chromatography of S-1 on phenyl-Sepharose is a potentially useful method for obtaining a purified myosin heavy-chain fragment with a high ATPase specific activity.",
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Hydrophobic interaction chromatography of myosin fragments : Potential use in purification. / Borejdo, Julian; Linder, Shoshana; Werber, Moshe M.

In: Archives of Biochemistry and Biophysics, Vol. 231, No. 1, 15.05.1984, p. 193-201.

Research output: Contribution to journalArticle

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