“Porin 31HL” in the Plasmalemma of Human Cells: A VDAC Discussed as Part of a Chloride Channel Complex in Normal and Cystic Fibrosis B-Lymphocyte Cell Lines

Thinnes, Friedrich P.; Babel, Dagmar; Heiden, Martin; Hein, A; Jürgens, Ludger; König, Ulrike; Hilschmann, Norbert

doi:10.1007/978-3-642-78936-6_27

Cited by 3 publications

(5 citation statements)

References 45 publications

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“…Concerning the physiological relevance of ATP-binding by VDAC, the proof given here of an ATP-binding site in human and bovine porin supports our proposal that plasmalemma integrated VDAC might form part of an ubiquitous chloride channel complex (Blatz and Magleby, 1983) which in patch clamping may figure as the outwardly rectifying depolarization induced chloride (ORDIC) channel 1990;1993;1994a;1994b;Thinnes 1992). In epithelial cells ORDIC is under the control of the cystic fibrosis transmembrane conductance regulator (CFTR) channel (Riordan et a/., 1989;Riordan 1993) and thus affected in cystic fibrosis (Egan etal., 1992;Gabriel et a/., 1993).…”

Section: Discussionsupporting

confidence: 77%

“…These observations on ATP or stilbene-disulfonate binding by mammalian porin, respectively, in combination with data from our and other laboratories on the expression of eukaryotic porin channels in the plasmalemma of different cells König et a/., 1991;Babel etal., 1991;Cole ef a/., 1992;Puchelle et a/., 1993;Janisch ef a/., 1993;Dermietzel ef a/., 1994) support our proposal that eukaryotic porin might be part of a chloride channel complex which in patch clamp experiments may figure as the outwardly rectifying depolarization induced chloride (ORDIC) channel in different cells (Thinnes ef a/., 1990;1993;1994a;1994b;Thinnes 1992).…”

Section: Introductionsupporting

confidence: 69%

“…Brought to you by | University of California Authenticated Download Date | 6/12/15 12:48 PM We had already pointed to a putative nucleotide binding site in mammalian porin and furthermore to the fact that VDAC of Neurospora crassa and Saccharomyces cerevisiae lack -at least corresponding -putative nucleotide binding sites (Thinnes et a/., 1994a). Concerning human porin the site: G Υ G F G goes from position 20 to 24 of the protein, forming part of the molecule which is likely to belong to its channel mouth (De Pinto et a/., 1991); the N-terminal region of the channel is expressed on the outer surface of the plasmalemma of different mammalian cells Cole et a/., 1992;Puchelle et a/., 1993;Janisch et a/., 1993;Dermietzel et a/., 1994) and on the cytosolic surface of mitochondria ( De Pinto et a/., 1991).…”

Section: Discussionmentioning

confidence: 96%

“…Arguments for plasmalemma integrated mammalian porin to make up part of the ORDIC channel complex have been broadly discussed and summarized in model form 1990;1993;1994a;1994b;Thinnes 1992).…”

Section: Discussionmentioning

confidence: 99%

“…We recently pointed to a putative nucleotide binding site of the human voltage-dependent anion channel (VDAC) "Porin 31HL" at position 20 to 24 of the molecule: G G F G and furthermore presented the first experimental data on reversible binding of adenosine triphosphate to channel active human porin by-using an affinity chromatography approach (Thinnes et a/., 1994a).…”

Section: Introductionmentioning

confidence: 99%

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Channel Active Mammalian Porin, Purified from Crude Membrane Fractions of Human B Lymphocytes and Bovine Skeletal Muscle, Reversibly Binds Adenosine Triphosphate (ATP)

Flörke¹,

Thinnes²,

Winkelbach³

et al. 1994

Biological Chemistry Hoppe-Seyler

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A new aspect of mammalian porin (mammalian VDAC = mammalian voltage-dependent anion channel) is presented: channel active VDAC binds adenosine triphosphate (ATP) in the absence of Ca2+. Channel active "Porin 31HL" or "Porin 31BM", enriched from crude membranes of human B lymphocytes or whole cell lysates of bovine skeletal muscle, respectively, was bound to a nine atoms spacer ATP-agarose at pH 7.4 or 5.0 and reeluted from the resin by 10 mM ATP disodium salt. Furthermore, channel active "Porin 31BM" was labelled by [32P]ATP in a 1:1 stoichiometric relation. Binding of ATP to human porin was confirmed by studying the interaction of the synthetic porin fragment Type-1/Ac-35, comprising the putative nucleotide binding site G Y G F G, with trinitrophenyl-ATP (TNT-ATP) by scanning fluorometry. Peptide/TNP-ATP complexes clearly show enhancement of fluorescence intensity and a spectral shift of the fluorescence maximum. In a control experiment, using a porin fragment lacking the putative nucleotide binding site, no change of fluorescence emission was observed. Further confirmation for ATP binding by human VDAC arose from an autoradiographic experimental approach: the porin fragment Type-1/Ac-35 could be labelled by [32P]ATP, while a second porin fragment ending immediately before the putative nucleotide binding site could not; nor could a synthetic non porin peptide.

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Section: Discussionsupporting

confidence: 77%

Section: Introductionsupporting

confidence: 69%

Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Channel Active Mammalian Porin, Purified from Crude Membrane Fractions of Human B Lymphocytes and Bovine Skeletal Muscle, Reversibly Binds Adenosine Triphosphate (ATP)

Flörke¹,

Thinnes²,

Winkelbach³

et al. 1994

Biological Chemistry Hoppe-Seyler

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New Findings Concerning Vertebrate Porin

Thinnes

Reymann

1997

Naturwissenschaften

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Eukaryotic porin can be considered to be a good candidate for forming the channel component of the protein complex which, depending on the approach used, may realize its expression either as the outwardly-rectifying depolarization-induced chloride channel or as the volume-sensitive organic osmolyte-anion channel. As a basis for this proposition, we point to a series of correspondences in properties between mammalian porin and the ORDIC channel complex. Specifically, mammalian porin is expressed in the plasmalemma of different cells and chloride channels can be blocked by anti-human porin antibodies in astrocytes and endothelial cells. There is an indication of colocalisation of human porin and the cystic fibrosis (CF) gene product, CFTR, in the apical region of epithelial cells. The primary structure of porin from a CF patient was found to be normal. Cytosol and amniotic fluid fractions influence the channel characteristics of mammalian porin. Channel-active mammalian porin binds ATP and the stilbene disulphonate grouping of the chloride channel inhibitor DIDS. Human porin in black membranes is a pathway for taurine, and biogenic polyamines reduce the voltage dependence of human porin. Assuming the relationship between human porin and the ORDIC channel/VSOAC complex, studies on plasmalemma-integrated human porin have a relevance for CF research. In addition, we refer to a case study on a child with encephalomyopathy in which porin could not be detected using monoclonal anti-human porin antibodies. Our studies were based on purified and sequenced human porin from different cells and from different cell compartments. In addition, we raised antibodies against mature human porin or synthetic parts of the molecule. This provided a firm foundation for our topochemical work with which we were able to establish the multi-topological expression of eukaryotic porin channels. The data are summarized and discussed.

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A case of serendipity*

Forrester

2007

Purinergic Signalling

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An account is given of how a sensitive bioassay system for measurement of the neurotransmitter acetylcholine serendipitously led to the identification of adenosine triphosphate (ATP) released in vitro from active skeletal muscle. Subsequent application of the identification procedures to exercising human muscle in vivo, cardiac muscle cells in vitro, and human erythrocytes exposed to hypoxia gave rise to the general concept of ATP as a molecule that could influence cell function from the extracellular direction. Mechanisms of ATP release from cells in terms of "trigger" events such as mechanical distortion of the membrane, depolarization of the membrane, and exposure to hypoxia are discussed. Potential therapeutic uses of extracellular ATP in cancer therapy, radiation therapy, and a possible influence upon aging are discussed. Possible roles (distant and local) of extracellular ATP released from muscle during whole body exercise are discussed.

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“Porin 31HL” in the Plasmalemma of Human Cells: A VDAC Discussed as Part of a Chloride Channel Complex in Normal and Cystic Fibrosis B-Lymphocyte Cell Lines

Cited by 3 publications

References 45 publications

Channel Active Mammalian Porin, Purified from Crude Membrane Fractions of Human B Lymphocytes and Bovine Skeletal Muscle, Reversibly Binds Adenosine Triphosphate (ATP)

Channel Active Mammalian Porin, Purified from Crude Membrane Fractions of Human B Lymphocytes and Bovine Skeletal Muscle, Reversibly Binds Adenosine Triphosphate (ATP)

New Findings Concerning Vertebrate Porin

A case of serendipity*

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