Highly sensitive and stable phosphatidylserine liposome aggregation assay for annexins

被引:24
作者
Lee, G
Pollard, HB
机构
[1] UNIFORMED SERV UNIV HLTH SCI, SCH MED, DEPT ANAT & CELL BIOL, BETHESDA, MD 20814 USA
[2] NIDDKD, CELL BIOL & GENET LAB, NIH, BETHESDA, MD 20892 USA
关键词
D O I
10.1006/abio.1997.2311
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Annexins are a gene family of Ca2+-dependent membrane binding proteins which interact specifically with acidic phospholipids. We describe here details of highly sensitive and precise assays for annexins I and V, utilizing turbidometric analysis of phosphatidylserine Liposome aggregation. In the case of annexin I, the new assay is 7-fold more sensitive than the conventional chromaffin granule aggregation assay in terms of threshold for detection and the rate of increase of initial absorbance is 15-fold greater. Annexin V, which binds but does not aggregate liposomes, can be assayed on the basis of inhibition of Ca2+-dependent liposome aggregation. Further comparative advantages of the assay include lower expense and increased shelf life of the liposome reagent.
引用
收藏
页码:160 / 164
页数:5
相关论文
共 30 条
[1]  
ANDREE HAM, 1990, J BIOL CHEM, V265, P4923
[2]   Similarity in calcium channel activity of annexin V and matrix vesicles in planar lipid bilayers [J].
Arispe, N ;
Rojas, E ;
Genge, BR ;
Wu, LNY ;
Wuthier, RE .
BIOPHYSICAL JOURNAL, 1996, 71 (04) :1764-1775
[3]   CHARACTERIZATION OF CA-2+-DEPENDENT PHOSPHOLIPID BINDING, VESICLE AGGREGATION AND MEMBRANE-FUSION BY ANNEXINS [J].
BLACKWOOD, RA ;
ERNST, JD .
BIOCHEMICAL JOURNAL, 1990, 266 (01) :195-200
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   Cyclic 3'-5'-adenosine monophosphate binds to annexin I and regulates calcium-dependent membrane aggregation and ion channel activity [J].
Cohen, BE ;
Lee, G ;
Arispe, N ;
Pollard, HB .
FEBS LETTERS, 1995, 377 (03) :444-450
[6]  
CREUTZ CE, 1978, J BIOL CHEM, V253, P2858
[7]   VESICLE AGGREGATION BY ANNEXIN-I - ROLE OF A SECONDARY MEMBRANE-BINDING SITE [J].
DELAFUENTE, M ;
PARRA, AV .
BIOCHEMISTRY, 1995, 34 (33) :10393-10399
[8]   AGGREGATION OF CHROMAFFIN GRANULES BY CALPACTIN AT MICROMOLAR LEVELS OF CALCIUM [J].
DRUST, DS ;
CREUTZ, CE .
NATURE, 1988, 331 (6151) :88-91
[9]   IDENTIFICATION OF A CELLULAR PROTEIN SUBSTRATE PHOSPHORYLATED BY THE AVIAN-SARCOMA VIRUS-TRANSFORMING GENE-PRODUCT [J].
ERIKSON, E ;
ERIKSON, RL .
CELL, 1980, 21 (03) :829-836
[10]  
FAVA RA, 1984, J BIOL CHEM, V259, P2636