Selective noncompetitive assimilation of bovine testicular beta-galactosidase and bovine liver beta-glucuronidase by generalized gangliosidosis fibroblasts.

Hieber, Virginia; Distler, Jack J.; Myerowitz, Rachel; Schmickel, Roy D.; Jourdian, George W.

doi:10.1172/jci109740

Cited by 9 publications

(5 citation statements)

References 25 publications

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“…Kinetic studies suggested that the endocytosis of acid hydrolases fulfills two requirements for a receptormediated, active transport process [6,[11][12][13][14][15][16][17][18][19]. First, selectivity was evident from rides (which were susceptible to degradation by alkaline phosphatase, a-mannosidase, and 0-N-acetylglucosaminidase) following endoglycosidase H treatment of high uptake a-N-acetylglucosaminidase.…”

Section: Evidence For Phosphorylmannose In the "Common Recognition Mamentioning

confidence: 99%

The Phosphomannosyl Recognition System for Intracellular and Intercellular Transport of Lysosomal Enzymes

Sly

1982

J of Cellular Biochemistry

364

152

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Section: Evidence For Phosphorylmannose In the "Common Recognition Mamentioning

confidence: 99%

The Phosphomannosyl Recognition System for Intracellular and Intercellular Transport of Lysosomal Enzymes

Sly

1982

J of Cellular Biochemistry

364

152

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“…proved to be acid hydrolases that were secreted into the culture medium by fibroblasts and were also present in body fluids and tissue extracts . The selectivity and saturability of the uptake system suggested a receptor-mediated uptake process (9,10,16,19,30) . I-cell disease fibroblasts, which were characterized by a deficiency for multiple lysosomal enzymes, were found to secrete enzymes into the extracellular fluid that were not susceptible to pinocytosis by normal fibroblasts .…”

mentioning

confidence: 99%

Chloroquine inhibits lysosomal enzyme pinocytosis and enhances lysosomal enzyme secretion by impairing receptor recycling.

González-Noriega¹,

Grubb²,

Talkad³

et al. 1980

The Journal of Cell Biology

715

279

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Adsorptive pinocytosis of acid hydrolases by fibroblasts depends on phosphomannosyl recognition markers on the enzymes and high-affinity pinocytosis receptors on the cell surface. In this study, ß-glucuronidase binding to the cell surface of attached fibroblasts was found to be saturable and inhibitable by mannose-6-phosphate (Man-6-P). Dissociation of cell-bound ß-glucuronidase occurred very slowly at neutral pH, but was greatly accelerated by lowering the pH below 6.0, or by exposure to Man-6-P. Comparison of the maximal cell surface binding and the observed rate of enzyme pinocytosis suggests that the pinocytosis receptors are replaced or reused about every 5 min . Enzyme pinocytosis was not affected by inhibition of new protein synthesis for several hours, suggesting a large pool of internal receptors and/or reuse of internalized receptors .Chloroquine treatment ofnormal human fibroblasts had three effects : (a) greatly enhanced secretion of newly synthesized acid hydrolases bearing the recognition marker for uptake, (b) depletion of enzyme-binding sites from the cell surface, and (c) inhibition of pinocytosis of exogenous enzyme . Only the third effect was seen in I-cell disease fibroblasts, which were also less sensitive than control cells to this effect .These observations are consistent with a model for transport of acid hydrolases that proposes that delivery of newly synthesized acid hydrolases to lysosomes requires the phosphomannosyl recognition marker on the enzymes, and intracellular receptors that segregate receptor-bound enzymes into vesicles for transport to lysosomes . This model explains how chloroquine, which raises intralysosomal pH, can disrupt both the intracellular pathway for newly synthesized acid hydrolases, and the one for uptake of exogenous enzyme by cell surface pinocytosis receptors .Adsorptive pinocytosis of lysosomal hydrolases was initially recognized as the uptake of "corrective factors" by enzyme-deficient fibroblasts (for review, see reference 14) . These corrective factors J . CELL BIOLOGY C The Rockefeller University Press -0021-9525/80/06/0839/14 $1.00 Volume 85 June 1980 839-852 proved to be acid hydrolases that were secreted into the culture medium by fibroblasts and were also present in body fluids and tissue extracts . The selectivity and saturability of the uptake system 839 on

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“…The more anionic, higher binding (14). Uptake of the pooled fractions by generalized gangliosidosis skin fibroblasts was performed as previously described (13). Both binding and uptake studies were performed with the subsaturating concentrations of &galactosidase indicated in the footnotes.…”

Section: Resultsmentioning

confidence: 99%

“…After 3 h of exposure the cells were scraped from the plates, dispersed, and washed in saline. Enzyme activity was determined after disruption by freezing and thawing (13). Cell protein was determined by the procedure of Lowry (15).…”

Section: Resultsmentioning

confidence: 99%

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Bovine testicular β-galactosidase: Purification of enzyme fractions that exhibit high affinity for phosphomannosyl receptors

Distler

Guo

Sahagian

et al. 1989

Analytical Biochemistry

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An improved method is described for the preparation of bovine testicular beta-galactosidase that allows the isolation of enzyme fractions that bind avidly to phosphomannosyl receptors. The procedure permits removal of a contaminating beta-hexosaminidase and yields nearly homogeneous beta-galactosidase. Enzyme eluted from DEAE-Sephacel was arbitrarily divided into pools that exhibited differing ability to bind phosphomannosyl receptors. A high binding fraction was rapidly assimilated by cultured cells and bound to both low and high molecular weight phosphomannosyl receptors. Carbohydrate analysis of the high binding fraction indicates an average content of one complex and one high mannose oligosaccharide chain per molecule and an average mannose 6-phosphate content of two residues per molecule. However, electrofocusing studies indicated that all the fractions were heterogeneous with respect to sialic acid and phosphate content. The purification procedure also provides highly purified beta-galactosidase suitable for removing beta-galactosidase residues from a variety of complex carbohydrates.

show abstract

Selective noncompetitive assimilation of bovine testicular beta-galactosidase and bovine liver beta-glucuronidase by generalized gangliosidosis fibroblasts.

Cited by 9 publications

References 25 publications

The Phosphomannosyl Recognition System for Intracellular and Intercellular Transport of Lysosomal Enzymes

The Phosphomannosyl Recognition System for Intracellular and Intercellular Transport of Lysosomal Enzymes

Chloroquine inhibits lysosomal enzyme pinocytosis and enhances lysosomal enzyme secretion by impairing receptor recycling.

Bovine testicular β-galactosidase: Purification of enzyme fractions that exhibit high affinity for phosphomannosyl receptors

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