Rapid micro-isolation of mammalian oxymyoglobin for biophysical studies

Dixie J. Goss, Lawrence J Parkhurst

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A rapid, single-step procedure is described for the isolation of mammalian oxymyoglobin for physical studies. The procedure employs a high-capacity, reusable, Hg-resin through which the myoglobin is filtered. The mammalian myoglobins for which sequences have been reported lack reactive cysteine. The column binds hemoglobin and other sulfhydryl-containing proteins, and retards othe proteins. The procedure requires less than twenty minutes for the isolation of oxymyoglobin from homogenized muscle tissue. Virtually no metmyoglobin is formed. The yields are 2-3 times greater than by other methods. The procedure can easily be scaled down for isolation of myoglobin from a few milligrams of muscle. Identical rate constants for oxygen association, dissociation, and CO association were measured by laser photolysis for beef heart myoglobin prepared by this procedure and that of Yamazaki et al. (Yamazaki, I., Yokota, K. and Shikama, K. (1964) J. Biol. Chem. 239, 4151). Gel electrophoresis indicates that the preparation is at least 95% pure.

Original languageEnglish (US)
Pages (from-to)315-322
Number of pages8
JournalJournal of Biochemical and Biophysical Methods
Volume3
Issue number6
DOIs
StatePublished - Dec 1980

Fingerprint

Myoglobin
Muscle
Metmyoglobin
Association reactions
Beef
Photolysis
Carbon Monoxide
Electrophoresis
Cysteine
Rate constants
Muscles
Hemoglobins
Proteins
Resins
Gels
Tissue
Oxygen
oxymyoglobin
Lasers

Keywords

  • mercuri-gel
  • micro-isolation
  • oxymyoglobin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

Cite this

Rapid micro-isolation of mammalian oxymyoglobin for biophysical studies. / Goss, Dixie J.; Parkhurst, Lawrence J.

In: Journal of Biochemical and Biophysical Methods, Vol. 3, No. 6, 12.1980, p. 315-322.

Research output: Contribution to journalArticle

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