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Studies on the nature of the subcellular localization of lactate dehydrogenase and glyceraldehyde‐3‐phosphate dehydrogenase in chicken skeletal muscle
Author(s) -
Melnick Ronald L.,
Hultin H. O.
Publication year - 1973
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040810202
Subject(s) - subcellular localization , homogenization (climate) , enzyme , differential centrifugation , biochemistry , glyceraldehyde 3 phosphate dehydrogenase , lactate dehydrogenase , organelle , dehydrogenase , biology , centrifugation , cytoplasm , biodiversity , ecology
The elucidation of the subcellular localization of enzymes by the classical technique of homogenization followed by differential centrifugation is limited in that it is difficult to determine the effect of the severe disruptive procedures on the normal relationship of the enzymes to their subcellular environment. Attempts have been made to study this problem under less severe limitations; one of the approaches used has been the use of pressure on whole muscle tissue to extract the cellular fluids. In this report we introduce the concept of “comparative extraction” for evaluation of results obtained by this procedure. By comparing the efflux of enzymes of similar solubility and similar size and shape, it is possible to determine the minimal amount of the less easily extractable enzyme which cannot be removed due to compartmentation or binding to cellular particulate structures. Using this concept of “comparative extraction,” we show in this report that at least 35% of the lactate dehydrogenase of chicken breast muscle is restricted in its removal. The data do not definitely resolve the problem of whether the restriction is due to compartmentation of the enzyme within subcellular organelles or binding to subcellular structures.