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Volumn 279, Issue 5356, 1998, Pages 1541-1544

Natural ligand of mouse CD1d1: Cellular glycosylphosphatidylinositol

Author keywords

[No Author keywords available]

Indexed keywords

CD1 ANTIGEN; GLYCOSYLPHOSPHATIDYLINOSITOL;

EID: 0032489652     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5356.1541     Document Type: Article
Times cited : (366)

References (35)
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    • note
    • 18 column (1,0 × 250 mm; Alltech) and eluted immediately by using a buffer B (0.05% TFA containing acetonitrile) gradient starting at 0% at 10 min to 37, 70, 90, and 100% at 73, 105, 115 and 120 min, respectively, at 50 μl/min.
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    • unpublished data
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    • note
    • 18 and cation-exchange column (1.0 × 250 mm; Alltech) by achieving 15, 60, and 100% buffer B at 15, 105, and 125 min, respectively, at 50 μl/min.
  • 19
    • 7144242122 scopus 로고    scopus 로고
    • note
    • Mass spectra were acquired on a Kratos analytical MALDI-4 mass spectrometer equipped with a curved-field reflectron and a nitrogen laser. About 0.3 to 1.0 μl of each fraction (30 to 40 μl), either directly or after concentrating to∼15 μl, was applied onto the sample probe. Matrix, saturated α-cyano-4-hydroxycinnamic acid in 45% ethanol containing 8.8% formic acid (∼300 nl), was then applied to the sample. A replicate of the sample was similarly spotted, except that, to enhance ion signal, 300 nl of saturated ammonium sulfate was added before matrix (20). Spectra were acquired in positive and negative modes as well as in linear and reflectron modes after application of a 20-kV accelerating voltage. Because different fragments form stable positive and negative ions, when derived from the same molecule, positive and negative spectra provide complementary structural information. The mass analyses reported here are within a mass accuracy of 0.5 dalton.
  • 23
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    • note
    • 18 RP-HPLC. Sample elution was initiated after the injection front returned to zero by increasing buffer B concentration to 37, 70, 90, and 100% at 63, 95, 105, and 110 min, respectively, at 50 μl/min.
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    • note
    • 3H]Mannose-labeled, sCD1d1 and Db-sol-associated ligands were separated from the heavy and light chains by Microcon-10 (Amicon) filtration to specifically monitor GPI-associated radioactivity.
  • 26
    • 7144244783 scopus 로고    scopus 로고
    • note
    • 3H]PI by Microcon-10 filtration, and radio-activity in the retained solution was measured.
  • 27
    • 7144246569 scopus 로고    scopus 로고
    • note
    • 3H]fucose and rarely to other sugars (33), there are about seven times as many mannoses and fucoses in the heavy chain as in GPI (77). Thus ∼65% of CD1d1 is occupied by GPI before accounting for losses incurred during the purification steps. Assuming 65 to 70% recovery of the ligand [based on peptide recoveries from class I molecules (15, 34)], then >90% of CD1d1 is occupied by GPI.
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    • note
    • b, re-spectiveiy; and A. K. Menon for helpful discussions. Supported by grants from NIH (SJ., S.P.B., and R.J.C.), the Juvenile Diabetes Foundation International (S.J.), the American Cancer Society (S.J.), and the National Research Council-NIH (R.R.B.).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.