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The localization of lysosomal enzymes in chromaffin tissue
Author(s) -
Smith A. D.,
Winkler H.
Publication year - 1966
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.1966.sp007859
Subject(s) - acid phosphatase , granule (geology) , biochemistry , differential centrifugation , enzyme , chemistry , cathepsin d , hydrolase , cathepsin , ribonuclease , chromatography , biology , rna , paleontology , gene
1. An homogenate of bovine adrenal medulla contains significant amounts of six acid hydrolases: acid ribonuclease, acid deoxyribonuclease, cathepsin, acid phosphatase, β‐glucuronidase and arylsulphatase. Most of the activity of each enzyme could be sedimented in the large‐granule fraction at 242,000 g ‐min. 2. Differential centrifugation indicated the presence of three populations of particles, which sedimented at slightly different rates; these are, in order of decreasing sedimentation rate, mitochondria, particles containing the acid hydrolases, and chromaffin granules. 3. The three types of particle could be separated by ultracentrifuging the large‐granule fraction in a sucrose density gradient. Most of the activity of each hydrolase was recovered in a layer intermediate between those formed by mitochondria and chromaffin granules. 4. The large‐granule fraction therefore contains particles which are defined by their enzyme content as lysosomes. 5. Highly purified chromaffin granules, containing less than 5% of the activity of each acid hydrolase, were obtained from the gradient.