Studies on Human Porin. VIII. Expression of “Porin 31HL” Channels in the Plasmalemma of the Acute-Lymphoblastic-Leukemia Cell Line KM3 as Revealed by Light- and Electron-Microscopy

Cole, T.; Awni, Lewa Adil; Nyakatura, Elke; Götz, H.; Walter, Götz; Thinnes, Friedrich P.; Hilschmann, Norbert

doi:10.1515/bchm3.1992.373.2.891

Cited by 31 publications

(13 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding was subsequently confirmed by a number of studies employing several cell lines or tissues (normal and cultured lymphocytes (Cole et al, 1992;Konig et al, 1991), epithelial cells (Puchelle et al, 1993), astrocytes (Dermietzel et al, 1994), and the post-synaptic membrane fraction from brain (Moon et al, 1999)), relaying on flow cytometry, as well as EM immunogold labeling and immunofluorescence techniques. Immunofluorescence showed a punctuate staining on the cell surface, compatible with a membrane subdomain localization of the protein.…”

Section: Vdac1 In the Plasma Membranementioning

confidence: 81%

VDAC, a multi-functional mitochondrial protein regulating cell life and death

Shoshan‐Barmatz

Pinto

Zweckstetter

et al. 2010

Molecular Aspects of Medicine

650

731

View full text Add to dashboard Cite

Section: Vdac1 In the Plasma Membranementioning

confidence: 81%

VDAC, a multi-functional mitochondrial protein regulating cell life and death

Shoshan‐Barmatz

Pinto

Zweckstetter

et al. 2010

Molecular Aspects of Medicine

650

731

View full text Add to dashboard Cite

“…VDAC is a channel protein that is mainly located on the mitochondrial outer membrane (27) but can also be found on the cell membrane of lymphocytes (28), epithelial cells (29), astrocytes (30), and the postsynaptic membrane fraction of the brain (31). In MASCOT analysis, GFMP-24 performed similarly to several protein candidates, whereas VDAC2 generated the highest similarity score (data not shown).…”

Section: Discussionmentioning

confidence: 96%

GHSC70 Is Involved in the Cellular Entry of Nervous Necrosis Virus

Chang

Chi

2015

J Virol

View full text Add to dashboard Cite

Nervous necrosis virus (NNV) is a devastating pathogen of cultured marine fish and has affected more than 40 fish species. NNV belongs to the betanodaviruses of Nodaviridae and is a nonenveloped icosahedral particle with 2 single-stranded positive-sense RNAs. To date, knowledge regarding NNV entry into the host cell remains limited, and no NNV-specific receptor protein has been published. Using grouper fin cell line GF-1 and purified NNV capsid protein in a virus overlay protein binding assay (VOPBA), grouper heat shock cognate protein 70 (GHSC70) and grouper voltage-dependent anion selective channel protein 2 (GVDAC2) were investigated as NNV receptor protein candidates. We cloned and sequenced the genes for GHSC70 and GVDAC2 and expressed them in Escherichia coli for antiserum preparation. Knockdown of the expression of GHSC70 and GVDAC2 genes with specific short interfering RNAs (siRNAs) significantly downregulated viral RNA expression in NNV-infected GF-1 cells. By performing an immunoprecipitation assay, we confirmed that GHSC70 interacted with NNV capsid protein, while VDAC2 did not. Immunofluorescence staining and flow cytometry analysis revealed the presence of the GHSC70 protein on the cell surface. After a blocking assay, we detected the NNV RNA2 levels after 1 h of adsorption to GF-1 cells; the level was significantly lower in the cells pretreated with the GHSC70 antiserum than in nontreated cells. Therefore, we suggest that GHSC70 participates in the NNV entry of GF-1 cells, likely functioning as an NNV receptor or coreceptor protein. IMPORTANCEFish nodavirus has caused mass mortality of more than 40 fish species worldwide and resulted in huge economic losses in the past 20 years. Among the four genotypes of fish nodaviruses, the red-spotted grouper nervous necrosis virus (RGNNV) genotype exhibits the widest host range. In our previous study, we developed monoclonal antibodies with high neutralizing efficiency against grouper NNV in GF-1 cells, indicating that NNV-specific receptor(s) may exist on the GF-1 cell membrane. However, no NNV receptor protein has been published. In this study, we found GHSC70 to be an NNV receptor (or coreceptor) candidate through VOBPA and provided several lines of evidence demonstrating that GHSC70 protein has a role in the NNV entry step of GF-1 cells. To the best of our knowledge, this is the first report identifying grouper HSC70 and its role in NNV entry into GF-1 cells.

show abstract

“…This antibody has been shown to recognize the N terminus of the HL-VDAC (19), which is highly homologous with our BR1-VDAC at the predicted amino acid level. Perhaps the N terminus of the HL-VDAC forms an extracytoplasmatic a-helix accessible for antibody detection (22). Fig.…”

Section: Gca Gtc Gtc Cgc Ccg Tat Act Gat Ctt Agc Aaa Tct Gcc Agg Gat Gtcmentioning

confidence: 99%

“…A large-conductance anion channel has been frequently described in astrocytes and we have stressed the strong similarity of the basic functional properties of this channel and that of mi-VDAC-type channels (see Introduction). By immunocytochemical studies, HL-VDAC has been localized in the plasma membrane of human lymphocytes (22). The same antibody (mAb2) was exploited in our blocking experiments.…”

Section: Gca Gtc Gtc Cgc Ccg Tat Act Gat Ctt Agc Aaa Tct Gcc Agg Gat Gtcmentioning

confidence: 99%

Cloning and in situ localization of a brain-derived porin that constitutes a large-conductance anion channel in astrocytic plasma membranes.

Dermietzel

Hwang

Buettner

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

137

View full text Add to dashboard Cite

We have cloned a protein from bovine brain, brain-derived voltage-dependent anion channel 1 (BR1-VDAC), that is identical to a recently sequenced plasmalemmal-bound porin from human lymphocytes. mRNA hybridization indicates that BR1-VDAC is widely distributed throughout nervous and nonnervous tissues. In situ localization substantiated that the BR1-VDAC is associated with the plasmalemma of astrocytes. A monoclonal antibody that recognizes the N terminus of the BR1-VDAC protein completely blocks an astrocytic high-conductance anion channel that has electrophysiological similarities with the mitochondrial VDAC. Since the high-conductance anion channel in astrocytes has been shown to respond to hypoosmotic solutions, its molecular identification provides the basis for a better understanding of volume regulation in brain tissue.Astrocytes are involved in the local homeostatic balancing of the interstitial cerebral fluid in brain. The correct condition of the interstitial cerebral fluid is crucial for normal functioning of the neuronal network. Astrocytes have a complement of neurotransmitter receptors and ion channels that form the backbone of this regulative capacity. Although physiological data on the diverse astrocytic receptor and channel types have been collected (1, 2), knowledge of their molecular composition is rare. Determining the molecular mechanisms involved in the homeostatic balancing of the interstitial cerebral fluid is, therefore, indispensible for an understanding of the properties of astrocytes and their concerted action in maintaining normal brain function. Among the various channel types electrophysiologically characterized in astrocytes (2, 3) is a large-conductance anion channel, described in cultured astrocytes (4-6) and cultured rat Schwann cells (7) and reported (8) to be present in astrocytes of intact optic nerves. Jalonen et al. (9) (30-34 kDa) and are considered to provide the pathway for the movement of nucleotides and various other substances into a variety of metabolic pathways (11-13). Compared with the large-conductance anion channel of astrocytes, they exhibit almost identical basic channel properties. (i) Singlechannel conductance is roughly 450 pS. (ii) They show ion selectivity for Cl-in the high-conductance state, which occurs at 0 mV transmembrane voltage. (iii) They respond symmetrically with respect to gating properties; i.e., they are in the high-conductance state at 0 mV and close to lowconductance states in response to application ofboth positive and negative potentials (14). The characteristic voltagedependent gating and the anion preference at low potentials have led to the term VDAC (11,12).VDACs in eukaryotes have been considered to be localized exclusively in mitochondria. Recently, however, an apparently plasmalemmal VDAC (plm-VDAC) protein highly homologous to mitochondrial porins has been sequenced from a human lymphocyte cell line by Edman degradation (15). Subsequently, another VDAC-like protein was found to copurify with the plasmalemmal-bound centra...

show abstract

Studies on Human Porin. VIII. Expression of “Porin 31HL” Channels in the Plasmalemma of the Acute-Lymphoblastic-Leukemia Cell Line KM3 as Revealed by Light- and Electron-Microscopy

Cited by 31 publications

References 2 publications

VDAC, a multi-functional mitochondrial protein regulating cell life and death

VDAC, a multi-functional mitochondrial protein regulating cell life and death

GHSC70 Is Involved in the Cellular Entry of Nervous Necrosis Virus

Cloning and in situ localization of a brain-derived porin that constitutes a large-conductance anion channel in astrocytic plasma membranes.

Contact Info

Product

Resources

About