Applications of Immobilized Adenosine Triphosphate in the Study of Myosin

Raphael Lamed, Avraham Oplatka*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Agarose-bound adenosine triphosphate (ATP) columns have been prepared which are capable of specifically binding active myosin and one-headed myosin at a high ionic strength as well as active heavy meromyosin (HMM) and HMM subfragment 1 (HMM S-1) at both high and low ionic strengths. The double-headed myosin and HMM were found to have a greater affinity for a column than one-headed myosin and HMM S-1, respectively. Removal of inactive material and preparation of fully active and more homogeneous species could thus be achieved. The binding of myosin and one-headed myosin required the presence of Ca2+ or Mg2+ ions in the millimolar range and was accompanied by the splitting of the bound ATP. Thus, binding in this case is not a reversible process and is intimately associated with the occurrence of a chemical process. One is therefore faced with a new type of affinity chromatography for which we suggest the term “dynamic affinity chromatography.” Bound ATP columns are currently used in our laboratory as a routine procedure for the preparation of purified active myosin species.

Original languageEnglish
Pages (from-to)3137-3142
Number of pages6
JournalBiochemistry
Volume13
Issue number15
DOIs
StatePublished - 1 Jul 1974
Externally publishedYes

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