Apparently Normal Extracellular Acidic Alpha-mannosidase in Fibroblast Cultures from Patients with Mannosidosis

Overview

Journal Am J Hum Genet

Publisher Cell Press

Specialty Genetics

Date 1982 Jan 1

PMID 7081212

Citations 1

Authors

C L DeFranco

J Charrow

L C HAHN

H L NADLER

Affiliations

Soon will be listed here.

Abstract

Fibroblasts from patients with mannosidosis, cultured in medium supplemented with fetal calf serum from which acidic alpha-mannosidase (alpha-D-mannoside mannohydrolase, E.C.3.2.1.24) has been removed, secreted a normal amount of apparently unaffected acidic alpha-mannosidase into fetal calf serum-free medium. Both the intracellular and extracellular acidic alpha-mannosidase activities were completely precipitated by antiserum to placenta alpha-mannosidase B. In contrast to the heat-lability of the intracellular acidic alpha-mannosidase and its low affinity for artificial mannoside substrate, the extracellular enzyme exhibited both normal thermostability and normal kinetics. Mixing experiments with the intercellular enzymes suggested that the decreased activity in the patients' fibroblasts is not the effect of an inhibitor or absence of an activator. However, incubation of the mannosidosis extracellular enzyme with either normal or patient cell lysate resulted in a partial loss of activity, whereas an additive value was observed with the normal extracellular enzyme. In contrast to normal culture medium, the medium from mannosidosis cell culture was unable to enhance the rate of reduction of intracellular radioactivity in mucolipidosis type II fibroblasts precultured in the presence of radiolabeled mannose. These findings suggest that the defect in mannosidosis is expressed only after the enzyme has been delivered to lysosomes and presumably undergone some form of processing there.

Citing Articles

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Raghavan S, Stuer G, Riviere L, Alroy J, Kolodny E J Inherit Metab Dis. 1988; 11(1):3-16.

PMID: 3128686 DOI: 10.1007/BF01800052.

References

Axen R, Porath J, Ernback S . Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967; 214(5095):1302-4. DOI: 10.1038/2141302a0. View

OCKERMAN P . Mannosidosis: isolation of oligosaccharide storage material from brain. J Pediatr. 1969; 75(3):360-5. DOI: 10.1016/s0022-3476(69)80259-3. View

Weber K, Osborn M . The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969; 244(16):4406-12. View

Marsh C, Gourlay G . Evidence for a non-lysosomal alpha-mannosidase in rat liver homogenates. Biochim Biophys Acta. 1971; 235(1):142-8. DOI: 10.1016/0005-2744(71)90041-6. View

Carroll M, DANCE N, Masson P, Robinson D, Winchester B . Human mannosidosis--the enzyme defect. Biochem Biophys Res Commun. 1972; 49(2):579-83. DOI: 10.1016/0006-291x(72)90450-0. View

Hickman S, Neufeld E . A hypothesis for I-cell disease: defective hydrolases that do not enter lysosomes. Biochem Biophys Res Commun. 1972; 49(4):992-9. DOI: 10.1016/0006-291x(72)90310-5. View

Poenaru L, DREYFUS J . Electrophoretic heterogeneity of human -mannosidase. Biochim Biophys Acta. 1973; 303(1):171-4. DOI: 10.1016/0005-2795(73)90158-x. View

Tsay G, Dawson G, Matalon R . Excretion of mannose-rich complex carbohydrates by a patient with alpha-mannosidase deficiency (mannosidosis). J Pediatr. 1974; 84(6):865-8. DOI: 10.1016/s0022-3476(74)80768-7. View

Taylor H, Thomas G, Aylsworth A, Stevenson R, REYNOLDS L . Mannosidosis: deficiency of a specific alpha-mannosidase component in cultured fibroblasts. Clin Chim Acta. 1975; 59(1):93-9. DOI: 10.1016/0009-8981(75)90223-5. View

10.

Neufeld E, Lim T, Shapiro L . Inherited disorders of lysosomal metabolism. Annu Rev Biochem. 1975; 44:357-76. DOI: 10.1146/annurev.bi.44.070175.002041. View

11.

Tsay G, Dawson G, Matalon R . Glycopeptide storage in skin fibroblasts cultured from a patient with alpha-mannosidase deficiency. J Clin Invest. 1975; 56(3):711-8. PMC: 301920. DOI: 10.1172/JCI108142. View

12.

Beaudet A, NICHOLS Jr B . Residual altered alpha-mannosidase in human mannosidosis. Biochem Biophys Res Commun. 1976; 68(1):292-8. DOI: 10.1016/0006-291x(76)90042-5. View

13.

Phillips N, Robinson D, Winchester B . Immunological characterization of human liver alpha-D-mannosidase. Biochem J. 1975; 151(3):469-75. PMC: 1172393. DOI: 10.1042/bj1510469a. View

14.

Booth C, Chen K, NADLER H . Mannosidosis: clinical and biochemical studies in a family of affected adolescents and adults. J Pediatr. 1976; 88(5):821-4. DOI: 10.1016/s0022-3476(76)81122-5. View

15.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

16.

Desnick R, Sharp H, Grabowski G, BRUNNING R, Quie P, Sung J . Mannosidosis: clinical, morphologic, immunologic, and biochemical studies. Pediatr Res. 1976; 10(12):985-96. DOI: 10.1203/00006450-197612000-00008. View

17.

Mersmann G, Buddecke E . Evidence for material from mannosidosis fibroblasts crossreacting with anti-acidic alpha-mannosidase antibodies. FEBS Lett. 1977; 73(1):123-6. View

18.

Hultberg B, Masson P . Normal extracellular excretion of acidic alpha-mannosidase activity by mannosidosis fibroblast cultures. Biochim Biophys Acta. 1977; 481(2):573-7. DOI: 10.1016/0005-2744(77)90289-3. View

19.

Tulsiani D, Opheim D, Touster O . Purification and characterization of alpha-D-mannosidase from rat liver golgi membranes. J Biol Chem. 1977; 252(10):3227-33. View

20.

Burton B, NADLER H . Quantitation of the enzymically deficient cross reacting material in GM1 gangliosidoses. Am J Hum Genet. 1977; 29(6):575-80. PMC: 1685510. View