Localization of Lactic Dehydrogenase Isozymes in Lysosomal Fraction of the Neutrophil of Normal Human Blood

John, Sosamma; Berger, Nathan A.; Bonner, Mary J.; Schultz, Julius

doi:10.1038/2151483b0

Cited by 10 publications

(5 citation statements)

References 7 publications

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“…In this case, however, it was possible to eliminate the explanation based on substrate permeability, by demonstrating latency of an exogenous lysosomal ,B-glucosidase (Lloyd, 1969). In human neutrophils lactate dehydrogenase is associated with lysosomes and is not latent (John, Berger, Bonner & Schultz, 1967). In reporting this John et al (1967) concluded that the enzyme, which is in most cells associated with the cytosol, in leucocytes adheres to the lysosomal membrane.…”

Section: Discussionmentioning

confidence: 97%

Studies on the permeability of rat liver lysosomes to carbohydrates

Lloyd

1969

Biochemical Journal

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1. The latency of nitrocatechol sulphatase activity was measured in rat liver lysosomes before and after preincubation in 0.25m solutions of 25 different carbohydrates. 2. Preincubation in disaccharides, hexitols, gluconate, glucuronate or lactate gave little or no rise in ;free' sulphatase activity, indicating that these compounds do not easily penetrate the lysosomal membrane, but incubation in monosaccharides or the lower glycitols caused a progressive loss of latency. 3. Rates of increase in ;free' activity were taken as an indication of rates of solute penetration into lysosomes and were correlated with the structure and molecular weight of each sugar. 4. Additional evidence for non-penetration of maltose was obtained by demonstrating that the latency of lysosomal alpha-glucosidase is independent of substrate concentration employed. 5. The results are discussed in the light of published data on the latency of lysosomal enzymes.

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Section: Discussionmentioning

confidence: 97%

Studies on the permeability of rat liver lysosomes to carbohydrates

Lloyd

1969

Biochemical Journal

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“…Acid phosphatase, measured with p-nitrophenyl phosphate, showed an intermediary distribution, in contrast with Vercauteren's data (27) and with our own. Schultz and co-workers have combined differential centrifugation with density gradient centrifugation in sucrose (25) or Ficoll (19) gradients to subfractionate granule fractions from human white blood cells. In these experiments, peroxidase was largely concentrated in a fraction of high density, together with considerable amounts of acid phosphatase and -glucuronidase; relatively high acid hydrolase activity occurred also in a fraction of lower density in which several mitochondrial enzymes were concentrated.…”

Section: Discussionmentioning

confidence: 99%

Resolution of Granules From Rabbit Heterophil Leukocytes Into Distinct Populations by Zonal Sedimentation

Baggiolini

Hirsch

Duve

1969

The Journal of Cell Biology

309

132

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Postnuclear supernates from homogenates of essentially pure rabbit heterophil leukocytes were fractionated by means of zonal differential centrifugation through a discontinuous sucrose gradient at various speeds. Three distinct groups of granules were characterized biochemically and morphologically. They were, in order of decreasing sedimentation coefficient: (a) Large, relatively dense granules, identified morphologically as the azurophil or primary granules, and containing essentially all of the myeloperoxidase activity of the preparations, about one-third of their lysozyme activity, and between 50 and 80% of their content in five acid hydrolases typically associated with lysosomes in other cells; (b) smaller, less dense granules, with the morphological appearance of the specific or secondary granules, and carrying most of the alkaline phosphatase and the remainder of the lysozyme activity of the preparations; (c) a second group of lysosome-like particles, associated with a morphologically heterogeneous fraction, and containing the remainder of the acid hydrolases, but little or no myeloperoxidase. When p-nitrophenyl phosphate was used instead of j-glycerophosphate for the assay of acid phosphatase, only small proportions of the total activity accompanied the two main lysosomal bands, and considerable activity was found in a zone slightly retarded with respect to the slowly moving band of acid hydrolases.

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“…The first attempt to isolate granule fractions from human PMNs was made by Schultz and co-workers (36,37). These authors have submitted granule preparations obtained by differential centrifugation to isopycnic equilibration in sucrose (36) and in Ficoll (37). They found that peroxidase was largely concentrated in a band with average density of 1.24, together with large amounts of ~-glucuronidase and acid phosphatase.…”

Section: Fractionation Of Human Pmn Homogenates In Other Laboratoriesmentioning

confidence: 99%

Biochemical and Morphological Characterization of Azurophil and Specific Granules of Human Neutrophilic Polymorphonuclear Leukocytes

Bretz¹,

Baggiolini²

1974

The Journal of Cell Biology

398

143

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Postnuclear supernates from homogenates of purified neutrophil polymorphonuclear leukocytes (PMNs) from human blood were fractionated by zonal sedimentation and isopycnic equilibration in sucrose gradients. The fractions were characterized biochemically by measuring protein content and the activities of eight enzymes. Selected fractions were further analyzed by electron microscopy. In both centrifugation systems, azurophil and specific granules could be resolved almost completely. Azurophil granules sediment three to four times faster than the specifics and have an average density of 1.23. They contain all the peroxidase of the cells, large portions of four lysosomal hydrolases, and about half of the total lysozyme, and therefore appear to be, in biochemical terms, very similar to the azurophil granules of rabbit PMNs. The specific granules, which have an average density of 1.19, contain the remaining half of the lysozyme but appear to be free of the other components of the azurophil granules, and of alkaline phosphatase. Isopycnic equilibration disclosed a minor lysosomal population, which strongly overlaps the specific granules, and made possible the identification of a membrane-fraction which is characterized by the presence of the thiol-sensitive acid 4-nitrophenyl phosphatase and of alkaline phosphatase.

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Localization of Lactic Dehydrogenase Isozymes in Lysosomal Fraction of the Neutrophil of Normal Human Blood

Cited by 10 publications

References 7 publications

Studies on the permeability of rat liver lysosomes to carbohydrates

Studies on the permeability of rat liver lysosomes to carbohydrates

Resolution of Granules From Rabbit Heterophil Leukocytes Into Distinct Populations by Zonal Sedimentation

Biochemical and Morphological Characterization of Azurophil and Specific Granules of Human Neutrophilic Polymorphonuclear Leukocytes

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