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Improved method for the isolation and preliminary characterization of human DAF (decay-accelerating factor).
DAF (decay-accelerating factor) is one of the integral membrane proteins of erythrocytes, and is considered to play an important role in the regulation of complement activation. The purification of DAF has been impeded by the difficulty in removing glycophorin. We devised an effective method for removing glycophorin. Through the limited trypsinization of stromata prior to the extraction of DAF, glycophorin was readily digested so that the DAF could be purified free of glycophorin by DEAE-Sephacel and Bio-Gel A 0.5 m chromatographies. On SDS-PAGE, DAF from trypsinized stromata showed the same mobility as that from native stromata: its molecular weight was estimated to be about 70 kDa. Amino acid analysis of DAF showed high contents of serine and glutamic acid. The amino-terminal sequence of DAF prepared by the present method, determined for the 29 residues, did not show significant homology with that of glycophorin.
PMID: 2428813 [PubMed - indexed for MEDLINE]
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Cited by 4 PubMed Central articles
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Decay-accelerating factor (DAF) in stool specimens as a marker of disease activity in patients with ulcerative colitis (UC).
Inaba T, Mizuno M, Ohya S, Kawada M, Uesu T, Nasu J, Takeuchi K, Nakagawa M, Okada H, Fujita T, et al.
Clin Exp Immunol. 1998 May; 112(2):237-41.
[Clin Exp Immunol. 1998]
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Identification of human erythrocyte blood group antigens on decay-accelerating factor (DAF) and an erythrocyte phenotype negative for DAF.
Telen MJ, Hall SE, Green AM, Moulds JJ, Rosse WF.
J Exp Med. 1988 Jun 1; 167(6):1993-8.
[J Exp Med. 1988]
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Role of human decay-accelerating factor in the evasion of Schistosoma mansoni from the complement-mediated killing in vitro.
Horta MF, Ramalho-Pinto FJ.
J Exp Med. 1991 Dec 1; 174(6):1399-406.
[J Exp Med. 1991]
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