المؤلفون: Samuel A. Lee, Summer M. Raines, Stella M. Bernardo, Karlett J. Parra, Hallie S. Rane, Jessica L. Binder

المصدر: Journal of Biological Chemistry. 288:6190-6201

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, Virulence Factors, ATPase, Quantitative Trait Loci, Mutant, Vacuole, Biology, Biochemistry, Membrane Biology, Catalytic Domain, Proton transport, Candida albicans, Molecular Biology, Ion Transport, Vacuolar lumen, Cell Biology, Hydrogen-Ion Concentration, biology.organism_classification, Cell biology, Vacuolar acidification, Biofilms, Vacuoles, biology.protein, Protons, Gene Deletion

الوصف: Vacuolar proton-translocating ATPase (V-ATPase) is a central regulator of cellular pH homeostasis, and inactivation of all V-ATPase function has been shown to prevent infectivity in Candida albicans. V-ATPase subunit a of the Vo domain (Voa) is present as two fungal isoforms: Stv1p (Golgi) and Vph1p (vacuole). To delineate the individual contribution of Stv1p and Vph1p to C. albicans physiology, we created stv1Δ/Δ and vph1Δ/Δ mutants and compared them to the corresponding reintegrant strains (stv1Δ/ΔR and vph1Δ/ΔR). V-ATPase activity, vacuolar physiology, and in vitro virulence-related phenotypes were unaffected in the stv1Δ/Δ mutant. The vph1Δ/Δ mutant exhibited defective V1Vo assembly and a 90% reduction in concanamycin A-sensitive ATPase activity and proton transport in purified vacuolar membranes, suggesting that the Vph1p isoform is essential for vacuolar V-ATPase activity in C. albicans. The vph1Δ/Δ cells also had abnormal endocytosis and vacuolar morphology and an alkalinized vacuolar lumen (pHvph1Δ/Δ = 6.8 versus pHvph1Δ/ΔR = 5.8) in both yeast cells and hyphae. Secreted protease and lipase activities were significantly reduced, and M199-induced filamentation was impaired in the vph1Δ/Δ mutant. However, the vph1Δ/Δ cells remained competent for filamentation induced by Spider media and YPD, 10% FCS, and biofilm formation and macrophage killing were unaffected in vitro. These studies suggest that different virulence mechanisms differentially rely on acidified vacuoles and that the loss of both vacuolar (Vph1p) and non-vacuolar (Stv1p) V-ATPase activity is necessary to affect in vitro virulence-related phenotypes. As a determinant of C. albicans pathogenesis, vacuolar pH alone may prove less critical than originally assumed.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ee650668e0cb76d859784f37c77678f5Test
https://doi.org/10.1074/jbc.m112.426197Test

المؤلفون: Jaekwon Lee, David J. Adle

المصدر: Journal of Biological Chemistry. 283:31460-31468

مصطلحات موضوعية: inorganic chemicals, Saccharomyces cerevisiae Proteins, Proteome, ATPase, Molecular Sequence Data, chemistry.chemical_element, Vacuole, Biology, Biochemistry, Gene Expression Regulation, Enzymologic, Substrate Specificity, Cytosol, Enzyme Stability, Amino Acid Sequence, Cation Transport Proteins, Molecular Biology, Adenosine Triphosphatases, Cadmium, Cell Biology, Endocytosis, Yeast, Transport protein, Membrane Transport, Structure, Function, and Biogenesis, chemistry, Vacuoles, Biophysics, biology.protein, Efflux, Copper, Intracellular

الوصف: Cadmium is a highly toxic environmental contaminant implicated in various diseases. Our previous data demonstrated that Pca1, a P1B-type ATPase, plays a critical role in cadmium resistance in yeast S. cerevisiae by extruding intracellular cadmium. This illustrates the first cadmium-specific efflux pump in eukaryotes. In response to cadmium, yeast cells rapidly enhance expression of Pca1 by a post-transcriptional mechanism. To gain mechanistic insights into the cadmium-dependent control of Pca1 expression, we have characterized the pathway for Pca1 turnover and the mechanism of cadmium sensing that leads to up-regulation of Pca1. Pca1 is a short-lived protein (t½ < 5 min) and is subject to ubiquitination when cells are growing in media lacking cadmium. Distinct from many plasma membrane transporters targeted to the vacuole for degradation via endocytosis, cells defective in this pathway did not stabilize Pca1. Rather, Pca1 turnover was dependent on the proteasome. These data suggest that, in the absence of cadmium, Pca1 is targeted for degradation before reaching the plasma membrane. Mapping of the N terminus of Pca1 identified a metal-responding degradation signal encompassing amino acids 250–350. Fusion of this domain to a stable protein demonstrated that it functions autonomously in a metal-responsive manner. Cadmium sensing by cysteine residues within this domain circumvents ubiquitination and degradation of Pca1. These data reveal a new mechanism for substrate-mediated control of P1B-type ATPase expression. Cells have likely evolved this mode of regulation for a rapid and specific cellular response to cadmium.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::349cdfffcbd5275021596a99fe3a1f2aTest
https://doi.org/10.1074/jbc.m806054200Test

المؤلفون: Gloria A. Martínez-Muñoz, Patricia M. Kane

المصدر: Journal of Biological Chemistry. 283:20309-20319

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, ATPase, Vacuole, Saccharomyces cerevisiae, Biochemistry, Cell membrane, Cytosol, medicine, Homeostasis, Enzyme Inhibitors, Molecular Biology, biology, Cell Membrane, Cell Biology, Hydrogen-Ion Concentration, Proton pump, Cell biology, Membrane Transport, Structure, Function, and Biogenesis, Protein Transport, medicine.anatomical_structure, Vacuolar acidification, Mutation, Vacuoles, biology.protein, Additions and Corrections, Macrolides, Protons

الوصف: Vacuolar proton-translocating ATPases (V-ATPases) play a central role in organelle acidification in all eukaryotic cells. To address the role of the yeast V-ATPase in vacuolar and cytosolic pH homeostasis, ratiometric pH-sensitive fluorophores specific for the vacuole or cytosol were introduced into wild-type cells and vma mutants, which lack V-ATPase subunits. Transiently glucose-deprived wild-type cells respond to glucose addition with vacuolar acidification and cytosolic alkalinization, and subsequent addition of K+ ion increases the pH of both the vacuole and cytosol. In contrast, glucose addition results in an increase in vacuolar pH in both vma mutants and wild-type cells treated with the V-ATPase inhibitor concanamycin A. Cytosolic pH homeostasis is also significantly perturbed in the vma mutants. Even at extracellular pH 5, conditions optimal for their growth, cytosolic pH was much lower, and response to glucose was smaller in the mutants. In plasma membrane fractions from the vma mutants, activity of the plasma membrane proton pump, Pma1p, was 65–75% lower than in fractions from wild-type cells. Immunofluorescence microscopy confirmed decreased levels of plasma membrane Pma1p and increased Pma1p at the vacuole and other compartments in the mutants. Pma1p was not mislocalized in concanamycin-treated cells, but a significant reduction in cytosolic pH under all conditions was still observed. We propose that short-term, V-ATPase activity is essential for both vacuolar acidification in response to glucose metabolism and for efficient cytosolic pH homeostasis, and long-term, V-ATPases are important for stable localization of Pma1p at the plasma membrane.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a68c8d134d1bb8f3c7c14231f33a19f2Test
https://doi.org/10.1074/jbc.m710470200Test

المؤلفون: Jie Qi, Michael Forgac

المصدر: Journal of Biological Chemistry. 282:24743-24751

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, ATPase, Mutant, Golgi Apparatus, Saccharomyces cerevisiae, Vacuole, Biology, Models, Biological, Biochemistry, Article, Catalysis, Dissociation (chemistry), Fungal Proteins, Proton transport, Protein Isoforms, V-ATPase, Enzyme Inhibitors, Molecular Biology, Adenosine Triphosphatases, Fungal protein, Wild type, Cell Biology, Cell biology, Glucose, Mutation, Vacuoles, Mutagenesis, Site-Directed, biology.protein, Macrolides

الوصف: One mechanism of regulating V-ATPase activity in vivo involves reversible dissociation into its component V(1) and V(0) domains, which in yeast occurs in response to glucose depletion. V-ATPase complexes containing the Vph1p isoform of subunit a (VCC) are targeted to the vacuole, and Stv1p-containing complexes (SCC) are targeted to the Golgi. Overexpression of Stv1p results in mistargeting of SCC to the vacuole. We have investigated the role of the a subunit isoform and cellular environment in controlling dissociation using vacuolar protein sorting (vps) mutants that accumulate proteins in either the prevacuolar compartment (PVC) (vps27Delta) or a post-Golgi compartment (PGC) (vps21Delta). Dissociation of both VCC and SCC depends upon cellular environment, with dissociation most complete in the vacuole and least complete in the PVC. The dependence of dissociation on V-ATPase activity was also investigated using both concanamycin and inactivating mutations. Concanamycin partly blocks dissociation of both VCC and SCC in all three compartments, with inhibition generally greater for SCC than VCC. The R735Q mutant of Vph1p results in loss of both ATPase and proton transport, whereas the R735K mutant lacks proton transport but has 10% of wild type ATPase activity. For VCC in the vacuole, dissociation is completely blocked for the R735Q but not the R735K mutant. Significant dissociation of VCC is observed for both mutants in the PVC and PGC, indicating that V-ATPase activity is not absolutely required for dissociation. Similar results were obtained for SCC, although dissociation of SCC is again generally more sensitive to activity than VCC. These results suggest that the cellular environment is important both in controlling in vivo dissociation of the V-ATPase and the dependence of this process on catalytic activity. Moreover, catalytic activity is not absolutely required for V-ATPase dissociation.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e28edc6b0032cd2e894fab0a03861be4Test
https://doi.org/10.1074/jbc.m700663200Test

المؤلفون: Elim Shao, Michael Forgac

المصدر: Journal of Biological Chemistry. 279:48663-48670

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, Time Factors, Immunoprecipitation, Protein subunit, ATPase, Blotting, Western, Genetic Vectors, Mutant, Saccharomyces cerevisiae, Vacuole, Biochemistry, Catalysis, Epitopes, Adenosine Triphosphate, Proton transport, Escherichia coli, V-ATPase, Molecular Biology, Dose-Response Relationship, Drug, biology, Wild type, Chloroquine, Cell Biology, Hydrogen-Ion Concentration, Protein Structure, Tertiary, Glucose, Quinacrine, Mutation, Vacuoles, biology.protein, Biophysics, Electrophoresis, Polyacrylamide Gel, Protons, Plasmids, Protein Binding

الوصف: The catalytic nucleotide binding subunit (subunit A) of the vacuolar proton-translocating ATPase (or V-ATPase) is homologous to the β-subunit of the F-ATPase but contains a 90-amino acid insert not present in the β-subunit, termed the nonhomologous region. We previously demonstrated that mutations in this region lead to changes in coupling of proton transport and ATPase activity and to inhibition of in vivo dissociation of the V-ATPase complex, an important regulatory mechanism (Shao, E., Nishi T., Kawasaki-Nishi, S., and Forgac, M. (2003) J. Biol. Chem. 278, 12985–12991). Measurement of the ATP dependence of coupling for the wild type and mutant proteins demonstrates that the coupling differences are observed at ATP concentrations up to 1 mm. A decrease in coupling efficiency is observed at higher ATP concentrations for the wild type and mutant V-ATPases. Immunoprecipitation of an epitope-tagged nonhomologous region from cell lysates indicates that this region is able to bind to the integral V0 domain in the absence of the remainder of the A subunit, an interaction confirmed by immunoprecipitation of V0. Interaction between the nonhomologous region and V0 is reduced upon incubation of cells in the absence of glucose, suggesting that the nonhomologous region may act as a trigger to activate in vivo dissociation. Immunoprecipitation suggests that the epitope tag on the nonhomologous region becomes less accessible upon glucose withdrawal, possibly due to binding to another cellular target. In vivo dissociation of the V-ATPase in response to glucose removal is also blocked by chloroquine, a weak base that neutralizes the acidic pH of the vacuole. The results suggest that the dependence of in vivo dissociation of the V-ATPase on catalytic activity may be due to neutralization of the yeast vacuole, which in turn blocks glucose-dependent dissociation.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b37a3951421db07460250ee2421951e0Test
https://doi.org/10.1074/jbc.m408278200Test

المؤلفون: Patricia M. Kane, Maria Sambade

المصدر: Journal of Biological Chemistry. 279:17361-17365

مصطلحات موضوعية: Glycerol, Vacuolar Proton-Translocating ATPases, Saccharomyces cerevisiae Proteins, Chromaffin Cells, Protein subunit, ATPase, Molecular Sequence Data, Mutant, Vacuole, Biology, medicine.disease_cause, Biochemistry, Epitopes, Open Reading Frames, Saccharomyces, Manduca, Centrifugation, Density Gradient, medicine, Animals, Amino Acid Sequence, Molecular Biology, Mutation, Sequence Homology, Amino Acid, Intracellular Membranes, Cell Biology, Hydrogen-Ion Concentration, biology.organism_classification, Yeast, Protein Structure, Tertiary, Open reading frame, Phenotype, Manduca sexta, Vacuoles, biology.protein, Cattle, Genome, Fungal, Protons, Plasmids

الوصف: The yeast cwh36Delta mutant was identified in a screen for yeast mutants exhibiting a Vma(-) phenotype suggestive of loss of vacuolar proton-translocating ATPase (V-ATPase) activity. The mutation disrupts two genes, CWH36 and a recently identified open reading frame on the opposite strand, YCL005W-A. We demonstrate that disruption of YCL005W-A is entirely responsible for the Vma(-) growth phenotype of the cwh36Delta mutant. YCL005W-A encodes a homolog of proteins associated with the Manduca sexta and bovine chromaffin granule V-ATPase. The functional significance of these proteins for V-ATPase activity had not been tested, but we show that the protein encoded by YCL005W-A, which we call Vma9p, is essential for V-ATPase activity in yeast. Vma9p is localized to the vacuole but fails to reach the vacuole in a mutant lacking one of the integral membrane subunits of the V-ATPase. Vma9p is associated with the yeast V-ATPase complex in vacuolar membranes, as demonstrated by co-immunoprecipitation with known V-ATPase subunits and glycerol gradient fractionation of solubilized vacuolar membranes. Based on this evidence, we propose that Vma9p is a genuine subunit of the yeast V-ATPase and that e subunits may be a functionally essential part of all eukaryotic V-ATPases.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::dfe5a78fbde77bfe63984b2f2e2cd0fdTest
https://doi.org/10.1074/jbc.m314104200Test

المؤلفون: Kevin J. Saliba, Stephanie Zissis, Stephen A. Ward, Kiaran Kirk, Richard J.W. Allen, Patrick G. Bray

المصدر: Journal of Biological Chemistry. 278:5605-5612

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, Erythrocytes, ATPase, Plasmodium falciparum, Vacuole, Biochemistry, chemistry.chemical_compound, Organelle, Animals, Humans, Malaria, Falciparum, Pyrophosphatases, Molecular Biology, Pyrophosphatase, Inorganic pyrophosphatase, biology, Hemozoin, Cell Biology, Hydrogen-Ion Concentration, biology.organism_classification, Inorganic Pyrophosphatase, Kinetics, Cytosol, chemistry, biology.protein, Digestion, Digestive System

الوصف: As it grows within the human erythrocyte, the malaria parasite, Plasmodium falciparum, ingests the erythrocyte cytosol, depositing it via an endocytotic feeding mechanism in the "digestive vacuole," a specialized acidic organelle. The digestive vacuole is the site of hemoglobin degradation, the storage site for hemozoin (an inert biocrystal of toxic heme), the site of action of many antimalarial drugs, and the site of proteins known to be involved in antimalarial drug resistance. The acidic pH of this organelle is thought to play a critical role in its various functions; however, the mechanisms by which the pH within the vacuole is maintained are not well understood. In this study, we have used a combination of techniques to demonstrate the presence on the P. falciparum digestive vacuole membrane of two discrete H(+) pumping mechanisms, both capable of acidifying the vacuole interior. One is a V-type H(+)-ATPase, sensitive to concanamycin A and bafilomycin A(1). The other is a H(+)-pyrophosphatase, which was inhibited by NaF and showed a partial dependence on K(+). The operation of the H(+)-pyrophosphatase was dependent on the presence of a Mg(2+)-pyrophosphate complex, and kinetic experiments gave results consistent with free pyrophosphate acting as an inhibitor of the protein. The presence of the combination of a H(+)-ATPase and a H(+)-pyrophosphatase on the P. falciparum digestive vacuole is similar to the situation in the acidic tonoplasts (vacuoles) of plant cells.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c82dbdb49bda63ab8be5a676a1edd489Test
https://doi.org/10.1074/jbc.m208648200Test

المؤلفون: Akira Kakizuka, Kiyoshi Inoue, Keiko Tanaka, Taeko Kobayashi

المصدر: Journal of Biological Chemistry. 277:47358-47365

مصطلحات موضوعية: Protein Folding, Time Factors, ATPase, Cell Cycle Proteins, Cell Separation, Vacuole, Endoplasmic Reticulum, PC12 Cells, Biochemistry, Mice, Cytosol, Valosin Containing Protein, Microscopy, Phase-Contrast, Endoplasmic Reticulum Chaperone BiP, Heat-Shock Proteins, Adenosine Triphosphatases, Neurons, Cell Death, biology, Cell Differentiation, Neurodegenerative Diseases, Flow Cytometry, AAA proteins, Cell biology, Plasmids, Protein Binding, Subcellular Fractions, Programmed cell death, DNA, Complementary, Valosin-containing protein, Green Fluorescent Proteins, Animals, Humans, RNA, Messenger, Molecular Biology, Cell Nucleus, Dose-Response Relationship, Drug, Endoplasmic reticulum, Cell Membrane, Cell Biology, Precipitin Tests, Molecular biology, Rats, Luminescent Proteins, Cytoplasm, Mutation, CCAAT-Enhancer-Binding Proteins, biology.protein, Unfolded protein response, Carrier Proteins, Transcription Factor CHOP, HeLa Cells, Molecular Chaperones, Transcription Factors

الوصف: Abnormal protein accumulation and cell death with cytoplasmic vacuoles are hallmarks of several neurodegenerative disorders. We previously identified p97/valosin-containing protein (VCP), an AAA ATPase with two conserved ATPase domains (D1 and D2), as an interacting partner of the Machado-Joseph disease (MJD) protein with expanded polyglutamines that causes Machado-Joseph disease. To reveal its pathophysiological roles in neuronal cells, we focused on its ATPase activity. We constructed and characterized PC12 cells expressing wild-type p97/VCP and p97(K524A), a D2 domain mutant. The expression level, localization, and complex formation of both proteins were indistinguishable, but the ATPase activity of p97(K524A) was much lower than that of the wild type. p97(K524A) induced cytoplasmic vacuoles that stained with an endoplasmic reticulum (ER) marker, and accumulation of polyubiquitinated proteins in the nuclear and membrane but not cytoplasmic fractions was observed, together with the elevation of ER stress markers. These results show that p97/VCP is essential for degrading membrane-associated ubiquitinated proteins and that profound deficits in its ATPase activity severely affect ER quality control, leading to abnormal ER expansion and cell death. Excessive accumulation of misfolded proteins may inactivate p97/VCP in several neurodegenerative disorders, eventually leading to the neurodegenerations.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::371665b795bf779501cb1a2d0c262202Test
https://doi.org/10.1074/jbc.m207783200Test

المؤلفون: A. Brett Mason, Thierry Ferreira, Carolyn W. Slayman, Kenneth E. Allen, Marc Pypaert

المصدر: Journal of Biological Chemistry. 277:21027-21040

مصطلحات موضوعية: biology, ATPase, Endoplasmic reticulum, Mutant, Cell Biology, Vacuole, Biochemistry, Yeast, Cell biology, biology.protein, Unfolded protein response, Molecular Biology, Secretory pathway, Biogenesis

الوصف: The yeast plasma-membrane H+-ATPase, encoded by PMA1, is delivered to the cell surface via the secretory pathway and has recently emerged as an excellent system for identifying quality control mechanisms along the pathway. In the present study, we have tracked the biogenesis of Pma1-G381A, a misfolded mutant form of the H+-ATPase. Although this mutant ATPase is arrested transiently in the peripheral endoplasmic reticulum, it does not become a substrate for endoplasmic reticulum-associated degradation nor does it appear to stimulate an unfolded protein response. Instead, Pma1-G381A accumulates in Kar2p-containing vesicular-tubular clusters that resemble those previously described in mammalian cells. Like their mammalian counterparts, the yeast vesicular-tubular clusters may correspond to specific exit ports from the endoplasmic reticulum, since Pma1-G381A eventually escapes from them (still in a misfolded, trypsin-sensitive form) to reach the plasma membrane. By comparison with wild-type ATPase, Pma1-G381A spends a short half-life at the plasma membrane before being removed and sent to the vacuole for degradation in a process that requires both End4p and Pep4p. Finally, in a separate set of experiments, Pma1-G381A was found to impose its phenotype on co-expressed wild-type ATPase, transiently retarding the wild-type protein in the ER and later stimulating its degradation in the vacuole. Both effects serve to lower the steady-state amount of wild-type ATPase in the plasma membrane and, thus, can explain the co-dominant genetic behavior of the G381A mutation. Taken together, the results of this study establish Pma1-G381A as a useful new probe for the yeast secretory system.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::b3199ceab8a53cf65fb413a8dfb4a711Test
https://doi.org/10.1074/jbc.m112281200Test

المؤلفون: Katherine Bowers, Tom H. Stevens, Shoko Kawasaki-Nishi, Tsuyoshi Nishi, Michael Forgac

المصدر: Journal of Biological Chemistry. 276:47411-47420

مصطلحات موضوعية: Vacuolar Proton-Translocating ATPases, Endosome, Recombinant Fusion Proteins, ATPase, Protein subunit, Amino Acid Motifs, Blotting, Western, Molecular Sequence Data, Golgi Apparatus, Vacuole, Biochemistry, chemistry.chemical_compound, symbols.namesake, Adenosine Triphosphate, ATP hydrolysis, Yeasts, Proton transport, Protein Isoforms, Amino Acid Sequence, Molecular Biology, Adenosine Triphosphatases, Sequence Homology, Amino Acid, biology, Hydrolysis, Cell Membrane, Cell Biology, Golgi apparatus, Precipitin Tests, Protein Structure, Tertiary, Glucose, Microscopy, Fluorescence, chemistry, Vacuoles, biology.protein, Biophysics, symbols, Protons, Adenosine triphosphate, Gene Deletion, Plasmids

الوصف: The 100-kDa "a" subunit of the vacuolar proton-translocating ATPase (V-ATPase) is encoded by two genes in yeast, VPH1 and STV1. The Vph1p-containing complex localizes to the vacuole, whereas the Stv1p-containing complex resides in some other intracellular compartment, suggesting that the a subunit contains information necessary for the correct targeting of the V-ATPase. We show that Stv1p localizes to a late Golgi compartment at steady state and cycles continuously via a prevacuolar endosome back to the Golgi. V-ATPase complexes containing Vph1p and Stv1p also differ in their assembly properties, coupling of proton transport to ATP hydrolysis, and dissociation in response to glucose depletion. To identify the regions of the a subunit that specify these different properties, chimeras were constructed containing the cytosolic amino-terminal domain of one isoform and the integral membrane, carboxyl-terminal domain from the other isoform. Like the Stv1p-containing complex, the V-ATPase complex containing the chimera with the amino-terminal domain of Stv1p localized to the Golgi and the complex did not dissociate in response to glucose depletion. Like the Vph1p-containing complex, the V-ATPase complex containing the chimera with the amino-terminal domain of Vph1p localized to the vacuole and the complex exhibited normal dissociation upon glucose withdrawal. Interestingly, the V-ATPase complex containing the chimera with the carboxyl-terminal domain of Vph1p exhibited a higher coupling of proton transport to ATP hydrolysis than the chimera containing the carboxyl-terminal domain of Stv1p. Our results suggest that whereas targeting and in vivo dissociation are controlled by sequences located in the amino-terminal domains of the subunit a isoforms, coupling efficiency is controlled by the carboxyl-terminal region.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0b6c5daccfe73631f61620796ed9d668Test
https://doi.org/10.1074/jbc.m108310200Test