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Decay accelerating factor of complement is anchored to cells by a C-terminal glycolipid

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00370a003· OSTI ID:6479444
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Membrane-associated decay accelerating factor (DAF) of human erythrocytes (E/sup hu/) was analyzed for a C-terminal glycolipid anchoring structure. Automated amino acid analysis of DAF following reductive radiomethylation revealed ethanolamine and glucosamine residues in proportions identical with those present in the E/sup hu/ acetylcholinesterase (AChE) anchor. Cleavage of radiomethylated 70-kilodalton (kDa) DAF with papain released the labeled ethanolamine and glucosamine and generated 61- and 55-kDa DAF products that retained all labeled Lys and labeled N-terminal Asp. Incubation of intact E/sup hu/ with phosphatidylinositol-specific phospholipase C (PI-PLC), which cleaves the anchors in trypanosome membrane form variant surface glycoproteins (mfVSGs) and murine thymocyte Thy-1 antigen, released 15% of the cell-associated DAF antigen. The released 67-kDa PI-PLC DAF derivative retained its ability to decay the classical C3 convertase C4b2a but was unable to membrane-incorporate and displayed physicochemical properties similar to urine DAF, a hydrophilic DAF form that can be isolated for urine. Nitrous acid deamination cleavage of E/sup hu/ DAF at glucosamine following labeling with the lipophilic photoreagent 3-(trifluoromethyl)-3-(m-(/sup 125/I)iodophenyl)diazirine ((/sup 125/I)TID) released the (/sup 125/I)TID label in a parallel fashion as from (/sup 125/I)TID-labeled AChE. Biosynthetic labeling of HeLa cells with (/sup 3/H) ethanolamine resulted in rapid /sup 3/H incorporation into both 48-kDa pro-DAF and 72-kDa mature epithelial cell DAF. The findings indicate that DAF and AChE are anchored in E/sup hu/ by the same or a similar glycolipid structure and that, like VSGs, this structure is incorporated into DAF early in DAF biosynthesis prior to processing of pro-DAF in the Golgi.
Research Organization:
Case Western Reserve Univ. School of Medicine, Cleveland, OH
OSTI ID:
6479444
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 25:22; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACID SEQUENCE
BETA DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CARBOXYLESTERASES
CELL CONSTITUENTS
CELL MEMBRANES
CHEMISTRY
CHOLINESTERASE
COMPLEMENT
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
ERYTHROCYTES
ESTERASES
GLUCOSAMINE
GLYCOLIPIDS
HELA CELLS
HEXOSAMINES
HEXOSES
HYDROLASES
IMMUNOASSAY
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIPIDS
MATERIALS
MEMBRANES
MOLECULAR STRUCTURE
MONOSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
SACCHARIDES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
UPTAKE