{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T06:44:05Z","timestamp":1779345845013,"version":"3.51.4"},"reference-count":25,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5391","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1998,11,6]]},"abstract":"<jats:p>\n            Adenosine triphosphate (ATP)\u2013sensitive potassium (K\n            <jats:sub>ATP<\/jats:sub>\n            ) channels couple cell metabolism to electrical activity. Phosphatidylinositol phosphates (PIPs) profoundly antagonized ATP inhibition of K\n            <jats:sub>ATP<\/jats:sub>\n            channels when applied to inside-out membrane patches. It is proposed that membrane-incorporated PIPs can bind to positive charges in the cytoplasmic region of the channel's K\n            <jats:sub>ir<\/jats:sub>\n            6.2 subunit, stabilizing the open state of the channel and antagonizing the inhibitory effect of ATP. The tremendous effect of PIPs on ATP sensitivity suggests that in vivo alterations of membrane PIP levels will have substantial effects on K\n            <jats:sub>ATP<\/jats:sub>\n            channel activity and hence on the gain of metabolism-excitation coupling.\n          <\/jats:p>","DOI":"10.1126\/science.282.5391.1138","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:44:47Z","timestamp":1027748687000},"page":"1138-1141","source":"Crossref","is-referenced-by-count":482,"title":["Membrane Phospholipid Control of Nucleotide Sensitivity of K\n            <sub>ATP<\/sub>\n            Channels"],"prefix":"10.1126","volume":"282","author":[{"given":"S.-L.","family":"Shyng","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Physiology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C. G.","family":"Nichols","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1146\/annurev.ne.11.030188.000525","volume":"11","author":"Ashcroft F. M.","year":"1988","unstructured":"Ashcroft F. M., Annu. Rev. Neurosci. 11, 97 (1988).","journal-title":"Annu. Rev. Neurosci."},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5239.1166"},{"key":"e_1_3_1_3_3","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1016\/S0896-6273(00)80124-5","volume":"16","author":"Inagaki N.","year":"1996","unstructured":"Inagaki N., et al., Neuron 16, 1011 (1996);","journal-title":"Neuron"},{"key":"e_1_3_1_3_4","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1085\/jgp.110.6.655","volume":"110","author":"Shyng S.-L.","year":"1997","unstructured":"Shyng S.-L., Nichols C. G., J. Gen. Physiol. 110, 655 (1997);","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_3_5","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1016\/S0896-6273(00)80321-9","volume":"18","author":"Clement J. P.","year":"1997","unstructured":"Clement J. P., et al., Neuron 18, 827 (1997).","journal-title":"Neuron"},{"key":"e_1_3_1_4_2","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1038\/387179a0","volume":"387","author":"Tucker S. J.","year":"1997","unstructured":"Tucker S. J., Gribble F. M., Zhao C., Trapp S., Ashcroft F. M., Nature 387, 179 (1997);","journal-title":"Nature"},{"key":"e_1_3_1_4_3","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1085\/jgp.110.2.141","volume":"110","author":"Shyng S.-L.","year":"1997","unstructured":"Shyng S.-L., Ferrigni T., Nichols C. G., J. Gen. Physiol. 110, 141 (1997).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"5388","DOI":"10.1074\/jbc.272.9.5388","volume":"272","author":"Fan Z.","year":"1997","unstructured":"Fan Z., Makielski J. C., J. Biol. Chem. 272, 5388 (1997);","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_5_3","doi-asserted-by":"publisher","DOI":"10.1126\/science.273.5277.956"},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1038\/35882","volume":"391","author":"Huang C. L.","year":"1998","unstructured":"Huang C. L., Feng S. Y., Hilgemann D. W., Nature 391, 803 (1998).","journal-title":"Nature"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1038\/305147a0","volume":"305","author":"Noma A.","year":"1983","unstructured":"Noma A., Nature 305, 147 (1983);","journal-title":"Nature"},{"key":"e_1_3_1_7_3","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1113\/jphysiol.1989.sp017869","volume":"419","author":"Lederer W. J.","year":"1989","unstructured":"Lederer W. J., Nichols C. G., J. Physiol. 419, 193 (1989).","journal-title":"J. Physiol."},{"key":"e_1_3_1_8_2","unstructured":"COSm6 cells were plated at \u223c2.5 \u00d7 10 5 cells per well (30-mm six-well dishes) and cultured in Dulbecco's modified Eagle's medium plus 10 mM glucose (DMEM-HG) with fetal calf serum (FCS 10%). Cells were transfected by incubation for 4 hours at 37 o C in DMEM medium plus 10% Nuserum; diethylaminoethyl-dextran (0.4 mg\/ml); 100 \u03bcM chloroquine; and 5 \u03bcg each of pCMV6b-K ir 6.2 pECE-SUR1 and pECE-GFP (green fluorescent protein) cDNA. Cells were incubated for 2 min in phosphate-buffered saline buffer containing DMSO (10%) and returned to DMEM-HG plus 10% FCS. Patch-clamp experiments were made at room temperature in a chamber (6) that allowed rapid change of the bathing solution. Micropipettes were filled with K-INT solution (below) electrode resistance \u223c0.5 to 1 M\u03a9). Patches were isolated from cells that fluoresced green under ultraviolet illumination and then voltage-clamped (Axopatch 1B Axon Foster City CA). The standard bath (intracellular) and pipette (extracellular) solution (K-INT) contained 140 mM KCl 10 mM K-Hepes and 1 mM K-EGTA (pH 7.3). PIP 2 was sonicated in ice for 30 min before application to inside-out patches. ATP sensitivity was assessed by brief exposures to ATP-containing solutions without PIP 2 (Fig. 1 A and B). Inward currents (shown as upward deflections) were measured at \u201350 mV filtered at 0.5 to 3 kHz digitized at 22 kHz (Neurocorder Neurodata NY ) and stored on videotape for offline analysis with the Microsoft Excel and Solver programs. Data are presented as the mean \u00b1 the standard error of the mean."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"1156","DOI":"10.1016\/S0006-3495(91)82151-8","volume":"60","author":"Cannell M. B.","year":"1991","unstructured":"Cannell M. B., Nichols C. G., Biophys. J. 60, 1156 (1991);","journal-title":"Biophys. J."},{"key":"e_1_3_1_9_3","unstructured":"; C. G. Nichols W. J. Lederer M. B. Cannell ibid. p.1164."},{"key":"e_1_3_1_10_2","first-page":"H1675","volume":"261","author":"Nichols C. G.","year":"1991","unstructured":"Nichols C. G., Lederer W. J., Am. J. Physiol. 261, H1675 (1991).","journal-title":"Am. J. Physiol."},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1085\/jgp.104.4.773","volume":"104","author":"Deutsch N.","year":"1994","unstructured":"Deutsch N., Matsuoka S., Weiss J. N., J. Gen. Physiol. 104, 773 (1994).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_12_2","unstructured":"S.-L. Shyng and C. G. Nichols data not shown."},{"key":"e_1_3_1_13_2","unstructured":"Single-letter abbreviations for the amino acid residues are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp; and Y Tyr."},{"key":"e_1_3_1_14_2","unstructured":"Mutant constructs were prepared by overlap extension at the junctions of the relevant residues by sequential polymerase chain reaction (PCR). Before transfection PCR products were subcloned into pCMV6b vector and sequenced. K ir 6.2-C was constructed by fusing a DNA fragment containing K ir 6.2 170-390 to pGEX2T vector. GST and K ir 6.2-C expressed in Escherichia coli were purified with glutathione-agarose beads and dialysis in K-INT solution (7)"},{"key":"e_1_3_1_15_2","unstructured":"Cells were incubated for 24 hours in culture medium containing 86 Rb Cl (1 \u03bcCi\/ml) 2 to 3 days after transfection. Before measurement of Rb efflux cells were incubated for 30 min at 25 o C in Krebs' Ringer solution with metabolic inhibitors [oligomycin (2.5 \u03bcg\/ml) plus 1 mM 2-deoxy- d -glucose]. At selected time points the solution was aspirated and replaced. The 86 Rb + in the aspirated solution was counted."},{"key":"e_1_3_1_16_2","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1126\/science.2122519","volume":"250","author":"Hoshi T.","year":"1991","unstructured":"Hoshi T., Zagotta W. N., Aldrich R. W., Science 250, 533 (1991).","journal-title":"Science"},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1146\/annurev.physiol.59.1.193","volume":"59","author":"Hilgemann D. W.","year":"1997","unstructured":"Hilgemann D. W., Annu. Rev. Physiol. 59, 193 (1997).","journal-title":"Annu. Rev. Physiol."},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/0014-5793(91)80259-6","volume":"279","author":"Findlay I.","year":"1991","unstructured":"Findlay I., Faivre J. F., FEBS Lett. 279, 95 (1991).","journal-title":"FEBS Lett."},{"key":"e_1_3_1_19_2","unstructured":"We are grateful to J. Shepard for technical assistance and to S. Seino and J. Nerbonne for K ir 6.2 and dissociated ventricular myocytes respectively. This work was supported by grant HL45742 from NIH (C.G.N.) an Established Investigatorship from the American Heart Association (C.G.N.) a Career Development Award from the American Diabetes Association (S.L.S.) and by the Washington University Diabetes Research Training Center."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.282.5391.1138","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T00:13:15Z","timestamp":1705104795000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.282.5391.1138"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,11,6]]},"references-count":25,"journal-issue":{"issue":"5391","published-print":{"date-parts":[[1998,11,6]]}},"alternative-id":["10.1126\/science.282.5391.1138"],"URL":"https:\/\/doi.org\/10.1126\/science.282.5391.1138","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,11,6]]}}}