Identification and properties of an ATPase in vacuolar membranes of Neurospora crassa
10.1128/jb.151.3.1326-1337.1982
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

Using a vacuolar preparation virtually free of contamination by other organelles, we isolated vacuolar membranes and demonstrated that they contain an ATPase. Sucrose density gradient profiles of vacuolar membranes show a single peak of ATPase activity at a density of 1.11 g/cm3. Comparison of this enzyme with the two well-studied proton-pumping ATPases of Neurospora plasma membranes and mitochondria shows that it is clearly distinct. The vacuolar membrane ATPase is insensitive to the inhibitors oligomycin, azide, and vanadate, but sensitive to N,N'-dicyclohexylcarbodiimide (Ki = 2 microM). It has a pH optimum of 7.5, requires a divalent cation (Mg2+ or Mn2+) for activity, and is remarkably unaffected (+/- 20%) by a number of monovalent cations, anions, and buffers. In its substrate affinity (Km for ATP = 0.2 mM), substrate preference (ATP greater than GTP, ITP greater than UTP greater than CTP), and loss of activity with repeated 1 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid washes, the vacuolar membrane ATPase resembles the F1F0 type of ATPase found in mitochondria and differs from the integral membrane type of ATPase in plasma membranes.

Bibliography

Bowman, E. J., & Bowman, B. J. (1982). Identification and properties of an ATPase in vacuolar membranes of Neurospora crassa. Journal of Bacteriology, 151(3), 1326–1337.

Authors 2
  1. E J Bowman (first)
  2. B J Bowman (additional)
References 51 Referenced 77
  1. Apps , D. K. , J. G. Pryde , R. Sutton , and J. H. Phillips . 1980 . Inhibition of adenosine triphosphatase, 5-hydroxytryptamine transport and proton translocation activities of resealed chromaffin-granule 'ghosts .' Biochem. J. 190 : 273 - 282 . (10.1042/bj1900273) / Biochem. J. / Inhibition of adenosine triphosphatase, 5-hydroxytryptamine transport and proton translocation activities of resealed chromaffin-granule 'ghosts by Apps D. K. (1980)
  2. Apps , D. K. , and G. A. Reid . 1977 . Adenosine triphosphatase and adenosine diphosphate/adenosine triphosphate isotope-exchange activities of the chromaffin-granule membrane . Biochem. J. 167 : 297 - 300 . (10.1042/bj1670297) / Biochem. J. / Adenosine triphosphatase and adenosine diphosphate/adenosine triphosphate isotope-exchange activities of the chromaffin-granule membrane by Apps D. K. (1977)
  3. Apps , D. K. , and G. Schatz . 1979 . An adenosine triphosphatase isolated from chromaffin-granule membranes is closely similar to Fl-adenosine triphosphatase of mitochondria . Eur. J. Biochem. 100 : 411 - 419 . (10.1111/j.1432-1033.1979.tb04184.x) / Eur. J. Biochem. / An adenosine triphosphatase isolated from chromaffin-granule membranes is closely similar to Fl-adenosine triphosphatase of mitochondria by Apps D. K. (1979)
  4. Boiler , T. , and H. Kende . 1979 . Hydrolytic enzymes in the central vacuole of the plant cell . Plant Physiol. 63 : 1123 - 1132 . (10.1104/pp.63.6.1123) / Plant Physiol. / Hydrolytic enzymes in the central vacuole of the plant cell by Boiler T. (1979)
  5. Bowman , B. J. , F. Blasco , and C. W. Slayman . 1981 . Purification and characterization of the plasma membrane ATPase of Neurospora crassa . J. Biol. Chem. 256 : 12343 - 12349 . (10.1016/S0021-9258(18)43277-2) / J. Biol. Chem. / Purification and characterization of the plasma membrane ATPase of Neurospora crassa by Bowman B. J. (1981)
  6. Bowman , B. J. , S. E. Mainzer , K. E. Alien , and C. W. Slayman . 1978 . Effects of inhibitors on the plasma membrane and mitochondrial adenosine triphosphatases of Neurospora crassa . Biochim. Biophys. Acta 512 : 13 - 28 . (10.1016/0005-2736(78)90214-6) / Biochim. Biophys. Acta / Effects of inhibitors on the plasma membrane and mitochondrial adenosine triphosphatases of Neurospora crassa by Bowman B. J. (1978)
  7. Bowman , B. J. , and C. W. Slayman . 1977 . Characterization of plasma membrane adenosine triphosphatase of Neurospora crassa . J. Biol. Chem. 252 : 3357 - 3363 . (10.1016/S0021-9258(17)40397-8) / J. Biol. Chem. / Characterization of plasma membrane adenosine triphosphatase of Neurospora crassa by Bowman B. J. (1977)
  8. Bowman , B. J. , and C. W. Slayman . 1979 . The effects of vanadate on the plasma membrane ATPase of Neurospora crassa . J. Biol. Chem. 254 : 2928 - 2934 . (10.1016/S0021-9258(17)30163-1) / J. Biol. Chem. / The effects of vanadate on the plasma membrane ATPase of Neurospora crassa by Bowman B. J. (1979)
  9. Bowman , E. J. , B. J. Bowman , and C. W. Slayman . 1981 . Isolation and characterization of plasma membranes from wild type Neurospora crassa . J. Biol. Chem. 256 : 12336 - 12342 . (10.1016/S0021-9258(18)43276-0) / J. Biol. Chem. / Isolation and characterization of plasma membranes from wild type Neurospora crassa by Bowman E. J. (1981)
  10. Burton , E. G. , and R. L. Metzenberg . 1974 . Properties of repressible alkaline phosphatase from wild type and a wall-less mutant of Neurospora crassa . J. Biol. Chem. 249 : 4679 - 4688 . (10.1016/S0021-9258(19)42374-0) / J. Biol. Chem. / Properties of repressible alkaline phosphatase from wild type and a wall-less mutant of Neurospora crassa by Burton E. G. (1974)
  11. Cramer , C. L. , L. E. Vaughn , and R. H. Davis . 1980 . Basic amino acids and inorganic polyphosphates in Neurospora crassa: independent regulation of vacuolar pools . J. Bacteriol. 142 : 945 - 952 . (10.1128/jb.142.3.945-952.1980) / J. Bacteriol. / Basic amino acids and inorganic polyphosphates in Neurospora crassa: independent regulation of vacuolar pools by Cramer C. L. (1980)
  12. d' Auzac , J. 1977 . ATPase membranaire de vacuoles Iysosomales: les Iutoids du latex d'Hevea brasiliensis . Phytochemistry 16 : 1881 - 1885 . (10.1016/0031-9422(77)80088-5) / Phytochemistry / ATPase membranaire de vacuoles Iysosomales: les Iutoids du latex d'Hevea brasiliensis by Auzac J. (1977)
  13. Davis , R. H. , B. J. Bowman , and R. L. Weiss . 1978 . Intracellular compartmentation and transport of metabolites . J. Supramol. Struct. 9 : 473 - 488 . (10.1002/jss.400090403) / J. Supramol. Struct. / Intracellular compartmentation and transport of metabolites by Davis R. H. (1978)
  14. De Duve , C. , and R. Watffaux . 1966 . Functions of Iysosomes. Annu. Rev. Physiol. 28 : 435 - 492 . (10.1146/annurev.ph.28.030166.002251) / Iysosomes. Annu. Rev. Physiol. / Functions of by De Duve C. (1966)
  15. Drams , C. , and R. L. Weiss . 1982 . Energetics of vacuolar compartmentation of arginine in Neurospora crassa . J. Bacteriol. 150 : 770 - 778 . (10.1128/jb.150.2.770-778.1982) / J. Bacteriol. / Energetics of vacuolar compartmentation of arginine in Neurospora crassa by Drams C. (1982)
  16. Dryer , R. L. , A. R. Tammes , and J. I. Routh . 1957 . The determination of phosphorus and phosphates with Nphenyl-p-phenylenediamine . J. Biol. Chem. 225 : 177 - 183 . (10.1016/S0021-9258(18)64920-8) / J. Biol. Chem. / The determination of phosphorus and phosphates with Nphenyl-p-phenylenediamine by Dryer R. L. (1957)
  17. Diirr , M. , K. Urech , T. Boiler , A. Wlemken , J. Schwencke , and M. Nagy . 1979 . Sequestration of arginine by polyphosphate in vacuoles of yeast (Saccharomyces cerevisiae) . Arch. Microbiol. 121 : 169 - 175 . (10.1007/BF00689982) / Arch. Microbiol. / Sequestration of arginine by polyphosphate in vacuoles of yeast (Saccharomyces cerevisiae) by Diirr M. (1979)
  18. Flngme , R. H. 1980 . The proton-translocating pumps of oxidative phosphorylation. Annu . Rev. Biochem. 49 : 1079 - 1113 . (10.1146/annurev.bi.49.070180.005243) / Rev. Biochem. / The proton-translocating pumps of oxidative phosphorylation. Annu by Flngme R. H. (1980)
  19. Halvorson , H. , and L. ElRiss . 1958 . The purification and properties of an a-glucosidase of Saccharomyces italicus Y1225 . Biochim. Biophys. Acta 30 : 28 - 40 . (10.1016/0006-3002(58)90237-3) / Biochim. Biophys. Acta / The purification and properties of an a-glucosidase of Saccharomyces italicus Y1225 by Halvorson H. (1958)
  20. Jackl , G. , and W. Sebald . 1975 . Identification of two products of mitochondrial protein synthesis associated with mitochondrial adenosine triphosphatase from Neurospora crassa . Eur. J. Biochem. 54 : 97 - 106 . (10.1111/j.1432-1033.1975.tb04118.x) / Eur. J. Biochem. / Identification of two products of mitochondrial protein synthesis associated with mitochondrial adenosine triphosphatase from Neurospora crassa by Jackl G. (1975)
  21. Kakinuma , Y. , Y. Ohsumi , and Y. Anraku . 1981 . Properties of H+-translocating adenosine triphosphatase in vacuolar membranes of Saccharomyces cerevisiae . J. Biol. Chem. 256 : 10859 - 10863 . (10.1016/S0021-9258(19)68523-6) / J. Biol. Chem. / Properties of H+-translocating adenosine triphosphatase in vacuolar membranes of Saccharomyces cerevisiae by Kakinuma Y. (1981)
  22. King , T. E. 1967 . Preparation of succinate dehydrogenase and reconstitution of succinate oxidase . Methods Enzymol. 10 : 322 - 331 . (10.1016/0076-6879(67)10061-X) / Methods Enzymol. / Preparation of succinate dehydrogenase and reconstitution of succinate oxidase by King T. E. (1967)
  23. Kirshner , N. 1962 . Uptake of catecholamines by a particulate fraction of the adrenal medulla . J. Biol. Chem. 237 : 2311 - 2317 . (10.1016/S0021-9258(19)63439-3) / J. Biol. Chem. / Uptake of catecholamines by a particulate fraction of the adrenal medulla by Kirshner N. (1962)
  24. Legerton , T. L. , and R. L. Weiss . 1979 . Mobilization of sequestered metabolites into degradative reactions by nutritional stress in Neurospora . J. Bacteriol. 138 : 909 - 914 . (10.1128/jb.138.3.909-914.1979) / J. Bacteriol. / Mobilization of sequestered metabolites into degradative reactions by nutritional stress in Neurospora by Legerton T. L. (1979)
  25. Lowry , 0. H. , N. J. Rosebrough , A. L. Farr , and R. J. Randall . 1951 . Protein measurement with the Folin phenol reagent . J. Biol. Chem. 193 : 265 - 275 . (10.1016/S0021-9258(19)52451-6) / J. Biol. Chem. / Protein measurement with the Folin phenol reagent by Lowry 0. (1951)
  26. Malner , S. E. , and C. W. Slayman . 1978 . Mitochondrial adenosine triphosphatase of wild-type and poky Neurospora crassa . J. Bacteriol. 133 : 584 - 592 . (10.1128/jb.133.2.584-592.1978) / J. Bacteriol. / Mitochondrial adenosine triphosphatase of wild-type and poky Neurospora crassa by Malner S. E. (1978)
  27. Marn , B. , M. Marin-Lanza , and E. Komor . 1981 . The protonmotive potential difference across the vacuo-lysosomal membrane of Hevea brasiliensis (rubber tree) and its modification by a membrane-bound adenosine triphosphatase . Biochem. J. 198 : 365 - 372 . (10.1042/bj1980365) / Biochem. J. / The protonmotive potential difference across the vacuo-lysosomal membrane of Hevea brasiliensis (rubber tree) and its modification by a membrane-bound adenosine triphosphatase by Marn B. (1981)
  28. Martnola , E. , U. Heck , T. Boder , A. Wlemken , and P. Matle . 1979 . Some properties of vacuoles isolated from Neurospora crassa slime variant . Arch. Microbiol. 120 : 31 - 34 . (10.1007/BF00413268) / Arch. Microbiol. / Some properties of vacuoles isolated from Neurospora crassa slime variant by Martnola E. (1979)
  29. Marty F. D. Branton and R. A. Leigh. 1980. Plant vacuoles p. 625-658. In N. E. Tolbert (ed.) The biochemistry of plants vol. 1. Academic Press Inc. New York. (10.1016/B978-0-12-675401-8.50021-X)
  30. Matlle , P. 1978 . Biochemistry and function of vacuoles. Annu . Rev. Plant Physiol. 29 : 193 - 213 . (10.1146/annurev.pp.29.060178.001205) / Rev. Plant Physiol. / Biochemistry and function of vacuoles. Annu by Matlle P. (1978)
  31. Nhhimura , M. , and H. Beevers . 1978 . Hydrolases in vacuoles from castor bean endosperm . Plant Physiol. 62 : 44 - 48 . (10.1104/pp.62.1.44) / Plant Physiol. / Hydrolases in vacuoles from castor bean endosperm by Nhhimura M. (1978)
  32. Njus , D. , J. Knoth , and M. Zallakian . 1981 . Proton-linked transport in chromaffin granules . Curr. Top. Bioenerg. 11 : 107 - 147 . / Curr. Top. Bioenerg. / Proton-linked transport in chromaffin granules by Njus D. (1981)
  33. Ohsumi , Y. , and Y. Anraku . 1981 . Active transport of basic amino acids driven by a proton motive force in vacuolar membrane vesicles of Saccharomyces cerevisiae . J. Biol. Chem. 256 : 2079 - 2082 . (10.1016/S0021-9258(19)69736-X) / J. Biol. Chem. / Active transport of basic amino acids driven by a proton motive force in vacuolar membrane vesicles of Saccharomyces cerevisiae by Ohsumi Y. (1981)
  34. Ophelm , D. J. , and 0. Touster . 1978 . Lysosomal a-Dmannosidase of rat liver. Purification and comparison with the Golgi and cytosolic a-D-mannosidases . J. Biol. Chem. 253 : 1017 - 1023 . (10.1016/S0021-9258(17)38105-X) / J. Biol. Chem. / Lysosomal a-Dmannosidase of rat liver. Purification and comparison with the Golgi and cytosolic a-D-mannosidases by Ophelm D. J. (1978)
  35. Pedersen , P. L. 1975 . Mitochondrial adenosine triphosphatase . J. Bioenerg. 6 : 243 - 275 . (10.1007/BF01648966) / J. Bioenerg. / Mitochondrial adenosine triphosphatase by Pedersen P. L. (1975)
  36. Pennington , R. J. 1961 . Biochemistry of dystrophic muscle . Biochem. J. 80 : 649 - 654 . / Biochem. J. / Biochemistry of dystrophic muscle by Pennington R. J. (1961)
  37. Reeves , J. P. , and T. Reames . 1981 . ATP stimulates amino acid accumulation by Iysosomes incubated with amino acid methyl esters. Evidence for a Iysosomal proton pump . J. Biol. Chem. 256 : 6047 - 6053 . (10.1016/S0021-9258(19)69126-X) / J. Biol. Chem. / ATP stimulates amino acid accumulation by Iysosomes incubated with amino acid methyl esters. Evidence for a Iysosomal proton pump by Reeves J. P. (1981)
  38. Scarborough , G. A. 1977 . Properties of the Neurospora crassa plasma membrane ATPase . Arch. Biochem. Biophys. 180 : 384 - 393 . (10.1016/0003-9861(77)90052-2) / Arch. Biochem. Biophys. / Properties of the Neurospora crassa plasma membrane ATPase by Scarborough G. A. (1977)
  39. Schneider , D. L. 1977 . Membranous localization and properties of ATPase of rat liver Iysosomes . J. Membr. Biol. 34 : 247 - 261 . (10.1007/BF01870302) / J. Membr. Biol. / Membranous localization and properties of ATPase of rat liver Iysosomes by Schneider D. L. (1977)
  40. Schneider , D. L. 1981 . ATP-dependent acidification of intact and disrupted Iysosomes . J. Biol. Chem. 256 : 3858 - 3864 . (10.1016/S0021-9258(19)69536-0) / J. Biol. Chem. / ATP-dependent acidification of intact and disrupted Iysosomes by Schneider D. L. (1981)
  41. Seargeant , L. E. , and R. A. Stinson . 1979 . Inhibition of human alkaline phosphatase by vanadate . Biochem. J. 181 : 247 - 250 . (10.1042/bj1810247) / Biochem. J. / Inhibition of human alkaline phosphatase by vanadate by Seargeant L. E. (1979)
  42. Sebald , W. , T. Graf , and H. B. Luklins . 1979 . The dicyclohexylcarbodiimide-binding protein of the mitochondrial ATPase complex from Neurospora crassa and Saccharomyces cerevisiae. Identification and isolation . Eur. J. Biochem. 93 : 587 - 599 . (10.1111/j.1432-1033.1979.tb12859.x) / Eur. J. Biochem. / The dicyclohexylcarbodiimide-binding protein of the mitochondrial ATPase complex from Neurospora crassa and Saccharomyces cerevisiae. Identification and isolation by Sebald W. (1979)
  43. van der Wilden , W. , and P. MatUle . 1978 . Isolation and characterization of yeast tonoplast fragments . Biochem. Physiol. Pflanz. 173 : 285 - 294 . (10.1016/S0015-3796(17)30496-1) / Biochem. Physiol. Pflanz. / Isolation and characterization of yeast tonoplast fragments by van der Wilden W. (1978)
  44. van Pilsum , J. F. , R. P. Martin , E. Kito , and J. Hess . 1956 . Determination of creatine, creatinine, arginine, guanidoacetic acid, guanidine, and methylguanidine in biological fluids . J. Biol. Chem. 222 : 225 - 236 . (10.1016/S0021-9258(19)50788-8) / J. Biol. Chem. / Determination of creatine, creatinine, arginine, guanidoacetic acid, guanidine, and methylguanidine in biological fluids by van Pilsum J. F. (1956)
  45. Vaughn , L. E. , and R. H. Davis . 1981 . Purification of vacuoles from Neurospora crassa . Mol. Cell. Biol. 1 : 797 - 806 . / Mol. Cell. Biol. / Purification of vacuoles from Neurospora crassa by Vaughn L. E. (1981)
  46. Vogel , H. J. 1964 . Distribution of Iysine pathways among fungi: evolutionary implications . Am. Nat. 98 : 435 - 446 . (10.1086/282338) / Am. Nat. / Distribution of Iysine pathways among fungi: evolutionary implications by Vogel H. J. (1964)
  47. Weiss , R. L. 1973 . Intracellular localization of ornithine and arginine pools in Neurospora . J. Biol. Chem. 248 : 5409 - 5413 . (10.1016/S0021-9258(19)43616-8) / J. Biol. Chem. / Intracellular localization of ornithine and arginine pools in Neurospora by Weiss R. L. (1973)
  48. Weiss , R. L. 1976 . Compartmentation and control of arginine metabolism in Neurospora . J. Bacteriol. 126 : 1173 - 1179 . (10.1128/jb.126.3.1173-1179.1976) / J. Bacteriol. / Compartmentation and control of arginine metabolism in Neurospora by Weiss R. L. (1976)
  49. Wiemken , A. , M. Schellenberg , and K. Urech . 1979 . Vacuoles: the sole compartment of digestive enzymes in yeast (Saccharomyces cerevisiae) . Arch. Microbiol. 123 : 23 - 35 . (10.1007/BF00403499) / Arch. Microbiol. / Vacuoles: the sole compartment of digestive enzymes in yeast (Saccharomyces cerevisiae) by Wiemken A. (1979)
  50. Wolf , H. U. , and L. Adolph . 1969 . Untersuchungen uber eine Mg 2+-(Ca 2+)-aktivierte ATPase aus Rinderhirnmikrosomen. Eur. J. Biochem. 8 : 68 - 74 . (10.1111/j.1432-1033.1969.tb00496.x) / Rinderhirnmikrosomen. Eur. J. Biochem. / Untersuchungen uber eine Mg 2+-(Ca 2+)-aktivierte ATPase aus by Wolf H. U. (1969)
  51. Yu , P. H. , M. R. Kula , and H. Tsai . 1976 . Isolation of specific protease inhibitors from Neurospora crassa . Can. J. Biochem. 54 : 699 - 703 . (10.1139/o76-100) / Can. J. Biochem. / Isolation of specific protease inhibitors from Neurospora crassa by Yu P. H. (1976)
Dates
Type When
Created 5 years, 8 months ago (Jan. 3, 2020, 12:37 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:28 p.m.)
Indexed 3 months, 2 weeks ago (May 20, 2025, 9:19 a.m.)
Issued 43 years ago (Sept. 1, 1982)
Published 43 years ago (Sept. 1, 1982)
Published Print 43 years ago (Sept. 1, 1982)
Funders 0

None

@article{Bowman_1982, title={Identification and properties of an ATPase in vacuolar membranes of Neurospora crassa}, volume={151}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.151.3.1326-1337.1982}, DOI={10.1128/jb.151.3.1326-1337.1982}, number={3}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Bowman, E J and Bowman, B J}, year={1982}, month=sep, pages={1326–1337} }