{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T11:10:55Z","timestamp":1768648255414,"version":"3.49.0"},"reference-count":19,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5403","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1999,2,5]]},"abstract":"<jats:p>\n            In T lymphocytes, a store-operated calcium ion (Ca\n            <jats:sup>2+<\/jats:sup>\n            ) entry mechanism termed the calcium release\u2013activated Ca\n            <jats:sup>2+<\/jats:sup>\n            channel (CRAC channel) underlies the sustained or oscillatory intracellular calcium concentration signal required for interleukin-2 gene expression and cell proliferation. The use of sodium ions as a current carrier enabled single-channel recordings of CRAC channels during activation, inactivation, and blockade of current in the presence of divalent cations. A large conductance of 36 to 40 picosiemens indicates that 100 to 400 CRAC channels are present in T lymphocytes.\n          <\/jats:p>","DOI":"10.1126\/science.283.5403.836","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:40:50Z","timestamp":1027762850000},"page":"836-839","source":"Crossref","is-referenced-by-count":118,"title":["Single-Channel Recording of a Store-Operated Ca\n            <sup>2+<\/sup>\n            Channel in Jurkat T Lymphocytes"],"prefix":"10.1126","volume":"283","author":[{"given":"Hubert H.","family":"Kerschbaum","sequence":"first","affiliation":[{"name":"Department of Animal Physiology, Institute of Zoology, University of Salzburg, A-5020 Salzburg, Austria."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Cahalan","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1016\/0143-4160(90)90016-N","volume":"11","author":"Putney J. W.","year":"1990","unstructured":"Putney J. W., Cell Calcium 11, 611 (1990);","journal-title":"Cell Calcium"},{"key":"e_1_3_1_2_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/bj3120001","volume":"312","author":"Berridge M. J.","year":"1995","unstructured":"Berridge M. J., Biochem. J. 312, 1 (1995).","journal-title":"Biochem. J."},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1146\/annurev.iy.13.040195.003203","volume":"13","author":"Lewis R. S.","year":"1995","unstructured":"Lewis R. S., Cahalan M. D., Annu. Rev. Immunol. 13, 623 (1995);","journal-title":"Annu. Rev. Immunol."},{"key":"e_1_3_1_3_3","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1016\/S0958-1669(97)80130-9","volume":"8","author":"Cahalan M. D.","year":"1997","unstructured":"Cahalan M. D., Chandy K. G., Curr. Opin. Biotechnol. 8, 749 (1997).","journal-title":"Curr. Opin. Biotechnol."},{"key":"e_1_3_1_4_2","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1091\/mbc.1.1.99","volume":"1","author":"Lewis R. S.","year":"1989","unstructured":"Lewis R. S., Cahalan M. D., Cell Regul. 1, 99 (1989);","journal-title":"Cell Regul."},{"key":"e_1_3_1_4_3","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1038\/355353a0","volume":"355","author":"Hoth M.","year":"1992","unstructured":"Hoth M., Penner R., Nature 355, 353 (1992).","journal-title":"Nature"},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"6295","DOI":"10.1073\/pnas.90.13.6295","volume":"90","author":"Zweifach A.","year":"1993","unstructured":"Zweifach A., Lewis R. S., Proc. Natl. Acad. Sci. U.S.A. 90, 6295 (1993).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1113\/jphysiol.1993.sp019681","volume":"465","author":"Hoth M.","year":"1993","unstructured":"Hoth M., Penner R., J. Physiol. (London) 465, 359 (1993).","journal-title":"J. Physiol. (London)"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/S0006-3495(96)79278-0","volume":"71","author":"Lepple-Wienhues A.","year":"1996","unstructured":"Lepple-Wienhues A., Cahalan M. D., Biophys. J. 71, 787 (1996).","journal-title":"Biophys. J."},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1085\/jgp.111.4.521","volume":"111","author":"Kerschbaum H. H.","year":"1998","unstructured":"Kerschbaum H. H., Cahalan M. D., J. Gen. Physiol. 111, 521 (1998).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_9_2","unstructured":"The human leukemic T cell line Jurkat E6-1 was cultured in RPMI 1640 with fetal calf serum (10%) 1 mM glutamine and 25 mM Hepes in a 5% CO 2 incubator at 37\u00b0C. Cells were plated on polylysine-coated glass cover slips immediately before whole-cell recording with Sylgard-coated pipettes fire-polished to a resistance of 2 to 5 megohms. (Sylgard was purchased from Dow Corning; all other chemicals were from Sigma.) All experiments were done at room temperature (\u223c20\u00b0C). Membrane currents were recorded with an EPC-9 patch clamp amplifier (HEKA Lambrecht Germany). Voltage stimuli lasting 200 ms were delivered from a holding potential of 0 mV every second using either voltage ramps in which voltage increased from \u2013120 to +40 mV or voltage steps from 0 to \u2013120 mV. Currents were sampled at 5 kHz during voltage ramps and at 25 kHz during voltage steps. Currents were digitally filtered off-line at 1 kHz. CRAC channels were opened by passive depletion of Ca 2+ stores with 12 mM BAPTA or by addition of 10 \u03bcM IP 3 with ionic conditions chosen to eliminate currents through K + or Cl \u2013 channels. Traces recorded before the activation of CRAC channels were used as a template to subtract leak currents. Internal solutions contained 128 mM Cs + aspartate 10 mM Cs + -Hepes (pH 7.2) 12 mM BAPTA 0.9 mM CaCl 2 and 3.16 mM MgCl 2 . Mg 2+ was omitted from Mg 2+ -free internal solution. Divalent-free external solution contained 150 mM Na + methane sulfonate 10 mM N -hydroxyethyl-ethylenediamine-triacetic acid (HEDTA) and 10 mM Hepes (pH 7.2). The external Ca 2+ solution contained 20 mM CaCl 2 in addition to 120 mM Na + methane sulfonate. We added 370 \u03bcM Mg 2+ to 150 mM Na + methane sulfonate with 10 mM HEDTA to yield a calculated 3 \u03bcM free Mg 2+ as computed with Maxchelator [D. Bers C. Patton R. Nuccitelli Eds. A Practical Guide to the Study of Ca 2+ in Living Cells vol. 40 of Methods in Cell Biology (Academic Press San Diego CA 1994)]."},{"key":"e_1_3_1_10_2","unstructured":"H. H. Kerschbaum and M. D. Cahalan unpublished data."},{"key":"e_1_3_1_11_2","unstructured":"Whole-cell Na + current densities after complete activation of CRAC channels averaged 153 \u00b1 44 pA\/pF with no Mg 2+ inside ( n = 45 cells) and 16 \u00b1 6 pA\/pF with internal Mg 2+ ( n = 10 cells)."},{"key":"e_1_3_1_12_2","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1085\/jgp.88.3.293","volume":"88","author":"Hess P.","year":"1986","unstructured":"Hess P., Lansman J. B., Tsien R. W., J. Gen. Physiol. 88, 293 (1986).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/309453a0"},{"key":"e_1_3_1_13_3","doi-asserted-by":"crossref","first-page":"7149","DOI":"10.1073\/pnas.82.20.7149","volume":"82","author":"McClesky E. W.","year":"1985","unstructured":"McClesky E. W., Almers W., Proc. Natl. Acad. Sci. U.S.A. 82, 7149 (1985);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_13_4","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1113\/jphysiol.1984.sp015351","volume":"353","author":"Almers W.","year":"1984","unstructured":"Almers W., McClesky E. W., Palade P. T., J. Physiol. (London) 353, 565 (1984).","journal-title":"J. Physiol. (London)"},{"key":"e_1_3_1_14_2","unstructured":"Conductance fluctuation analysis is likely to have underestimated the single-channel conductance and overestimated the number of CRAC channels per cell because the P o value of single channels is very high as soon as the CRAC channels begin to conduct. In stationary noise analysis the single-channel conductance \u03b3 can be estimated by measuring the variance and mean conductance values (\u03c3 g 2 and G ) and then scaling according to the equation \u03b3 = \u03c3 g 2 \/[ G (1 \u2013 P o )]. Using the value of P o = 0.94 determined here a previous noise estimate of the single-channel Na + conductance of 2.6 pS can be corrected resulting in a revised estimate of 43 pS in good agreement with our direct single-channel measurements (6). If the same P o value also applies to high-divalent conditions the variance and mean values of conductance estimated in (4) would correspond to a single-channel Ca 2+ conductance of \u223c400 fS. High-frequency block and unbinding by divalent ions above 1 kHz may also have lowered the variance and hence the single-channel Ca 2+ conductance estimated by noise analysis. An alternative approach to estimate the Ca 2+ conductance through a single CRAC channel is to divide the single-channel Na + conductance of 36 to 40 pS by the ratio of macroscopic Na + to Ca 2+ currents when divalents are withdrawn and then Ca 2+ reapplied a ratio that averaged 25 (7). This approach assumes a constant number of open channels immediately before and after addition of Ca 2+ and yields an estimate of 1.6 pS for the CRAC channel carrying Ca 2+ at a concentration of 20 mM."},{"key":"e_1_3_1_15_2","unstructured":"We thank L. Forrest for assistance in cell culture. Supported by NIH grants NS-14609 and GM-41514."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.283.5403.836","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T03:08:58Z","timestamp":1705115338000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.283.5403.836"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,2,5]]},"references-count":19,"journal-issue":{"issue":"5403","published-print":{"date-parts":[[1999,2,5]]}},"alternative-id":["10.1126\/science.283.5403.836"],"URL":"https:\/\/doi.org\/10.1126\/science.283.5403.836","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,2,5]]}}}