{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T19:35:49Z","timestamp":1791056149746,"version":"4.1.0"},"reference-count":35,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5538","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2001,9,21]]},"abstract":"<jats:p>SLAP-130\/Fyb (SLP-76\u2013associated phosphoprotein or Fyn-binding protein; also known as Fyb\/Slap) is a hematopoietic-specific adapter, which associates with and modulates function of SH2-containing leukocyte phosphoprotein of 76 kilodaltons (SLP-76). T cells from mice lacking SLAP-130\/Fyb show markedly impaired proliferation following CD3 engagement. In addition, the T cell receptor (TCR) in SLAP-130\/Fyb mutant cells fails to enhance integrin-dependent adhesion. Although TCR-induced actin polymerization is normal, TCR-stimulated clustering of the integrin LFA-1 is defective in SLAP-130\/Fyb\u2013deficient cells. These data indicate that SLAP-130\/Fyb is important for coupling TCR-mediated actin cytoskeletal rearrangement with activation of integrin function, and for T cells to respond fully to activating signals.<\/jats:p>","DOI":"10.1126\/science.1063486","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:39:52Z","timestamp":1027748392000},"page":"2263-2265","source":"Crossref","is-referenced-by-count":268,"title":["Coupling of the TCR to Integrin Activation by SLAP-130\/Fyb"],"prefix":"10.1126","volume":"293","author":[{"given":"Erik J.","family":"Peterson","sequence":"first","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"},{"name":"Medicine and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melody L.","family":"Woods","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Center for Immunology, Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sally A.","family":"Dmowski","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geo","family":"Derimanov","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"},{"name":"Medicine and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martha S.","family":"Jordan","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer N.","family":"Wu","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peggy S.","family":"Myung","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qing-Hua","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan T.","family":"Pribila","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Center for Immunology, Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce D.","family":"Freedman","sequence":"additional","affiliation":[{"name":"Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoji","family":"Shimizu","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, Center for Immunology, Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gary A.","family":"Koretzky","sequence":"additional","affiliation":[{"name":"The Abramson Family Cancer Research Institute, Departments of"},{"name":"Pathology and Laboratory Medicine, School of Medicine, and the"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/S0079-6107(98)00050-9","volume":"71","author":"Samelson L. 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The targeting vector was electroporated into R1 embryonic stem cells (129\/SVJ). G418-resistant embryonic stem cells were injected into day-4 blastocysts and implanted in pseudopregnant mice (InGenious Targeting Laboratory Stony Brook NY). Resulting chimeric mice were bred with wild-type B6 to generate animals heterozygous for the targeted allele (13)."},{"key":"e_1_3_1_11_2","unstructured":"For T cell purification T cells were negatively selected from red-cell depleted splenocytes using Biomag Sheep anti-FITC particles (Polysciences) after staining with FITC-conjugated anti-B220 anti-Mac-1 (Caltag each mAb 10 \u03bcl\/spleen) and anti-DX-5 (Pharmingen 5 \u03bcl\/spleen). Lymph node cells were tumbled with anti-B220 coupled magnetic beads (Dynal). After purification cell preparations were 88 to 95% CD3-positive."},{"key":"e_1_3_1_12_2","unstructured":"For assessment of LFA-1 clustering purified splenic T cells were incubated with 2C11 (1 \u03bcg\/10 6 cells) on ice for 30 min washed then cross-linked with goat anti-hamster Ig (0.5 \u03bcg\/10 6 cells) on ice for 30 min. After further washing cells were stimulated in 37\u00b0C PBS for 10 min fixed in 4% formaldehyde stained with anti-CD11a (25 \u03bcg\/ml Pharmingen) and plated on poly- L -lysine coated Labtek chamber slides. For each experiment a minimum of 50 cells from each condition were imaged by confocal microscopy and analyzed for LFA-1 staining pattern (33). Cells were divided into quadrants fluorescence intensity in each quadrant was determined and standard deviations (SD) between quadrant fluorescence intensities were calculated. Cells showing SD greater than 0.1 were \u201cpolarized \u201d while cells with SD of 0.099 or less were \u201cnon-polarized.\u201d"},{"key":"e_1_3_1_13_2","unstructured":"For actin and LFA-1 capping studies antibodies were conjugated with Dynal M450 beads per manufacturer instructions. Cells and beads were co-incubated at a 1:1 ratio for 20 min (RT) and allowed to settle on poly-L-lysine coated glass coverslips. Cells were fixed with 4% paraformaldehyde for 20 min then blocked with 1:50 normal rat serum in 1% BSA\/PBS for 60 min. Samples were stained with either FITC-conjugated anti-mouse CD11a (LFA-1\u03b1 chain 50 \u03bcg\/ml Pharmingen) mAb or with rhodamine phalloidin (300 nM) for 30 min washed three times with PBS and mounted on microscopy slides. Samples were visualized on a Nikon E-800 microscope equipped with CCD camera using fluorescent light with 480\/20 nm excitation and 535\/25 emission filters. In a blinded fashion 50 randomly selected cell-bead conjugates per sample were scored as either negative or positive for the presence of an actin or a LFA-1 cap at the cell-bead contact site."},{"key":"e_1_3_1_14_2","unstructured":"Supplemental material is available on Science Online at www.sciencemag.org\/cgi\/content\/full\/293\/5538\/2263\/DC1."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1084\/jem.190.7.943","volume":"190","author":"Freedman B. D.","year":"1999","unstructured":"Freedman B. D., Liu Q. H., Somersan S., Kotlikoff M. I., Punt J. A., J. Exp. Med. 190, 943 (1999).","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_16_2","unstructured":"E. J. Peterson Q. Liu B. Freedman M. Woods S. Dmowski Y. Shimizu G. A. Koretzky unpublished observations."},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","first-page":"4579","DOI":"10.4049\/jimmunol.144.12.4579","volume":"144","author":"Van Seventer G. 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