The ear of alpha-adaptin interacts with the COOH-terminal domain of the Eps15 protein

被引:165
作者
Benmerah, A
Begue, B
DautryVarsat, A
CerfBensussan, N
机构
[1] HOP NECKER ENFANTS MALAD,INSERM,U429,F-75743 PARIS 15,FRANCE
[2] INST PASTEUR,CNRS,URA 1960,UNITE BIOL INTERACT CELLULAIRES,F-75724 PARIS 15,FRANCE
关键词
D O I
10.1074/jbc.271.20.12111
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The role of Eps15 in clathrin-mediated endocytosis is supported by two observations. First, it interacts specifically and constitutively with the plasma membrane adaptor AP-2. Second, its NH2 terminus shows significant homology to the NH2 terminus of yeast End3p, necessary for endocytosis of Lu-factor. To gain further insight into the role of Eps15-AP-2 association, we have now delineated their sites of interactions. AP-2 binds to a domain of 72 amino acids (767-739) present in the COOH terminus of Fps15. This domain contains 4 of the 15 DPF repeats characteristic of the COOH-terminal domain of Eps15 and shares no homology with known proteins, including the related Fps15r protein. Precipitation of proteolytic fragments of AP-2 with Eps15-derived fusion proteins containing the binding site for AP-2 showed that Eps15 binds specifically to a 40-kDa fragment corresponding to the ear of alpha-adaptin, a result confirmed by precipitation of Eps15 by alpha-adaptin-derived fusion proteins. Our data indicate that this specific part of AP-2 binds to a cellular component and provide the tools for investigating the function(s) of the association between AP-2 and Eps15.
引用
收藏
页码:12111 / 12116
页数:6
相关论文
共 29 条
[11]   CLATHRIN AND ASSOCIATED ASSEMBLY AND DISASSEMBLY PROTEINS [J].
KEEN, JH .
ANNUAL REVIEW OF BIOCHEMISTRY, 1990, 59 :415-438
[12]  
KIRCHHAUSEN T, 1991, J BIOL CHEM, V266, P11153
[13]   STRUCTURAL AND FUNCTIONAL DIVISION INTO 2 DOMAINS OF THE LARGE (100-KDA TO 115-KDA) CHAINS OF THE CLATHRIN-ASSOCIATED PROTEIN COMPLEX AP-2 [J].
KIRCHHAUSEN, T ;
NATHANSON, KL ;
MATSUI, W ;
VAISBERG, A ;
CHOW, EP ;
BURNE, C ;
KEEN, JH ;
DAVIS, AE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (08) :2612-2616
[14]  
LINDNER R, 1992, J BIOL CHEM, V267, P16567
[15]  
NATKE IS, 1992, CELL, V68, P899
[16]   INTERACTION OF TYROSINE-BASED SORTING SIGNALS WITH CLATHRIN-ASSOCIATED PROTEINS [J].
OHNO, H ;
STEWART, J ;
FOURNIER, MC ;
BOSSHART, H ;
RHEE, I ;
MIYATAKE, S ;
SAITO, T ;
GALLUSSER, A ;
KIRCHHAUSEN, T ;
BONIFACINO, JS .
SCIENCE, 1995, 269 (5232) :1872-1875
[17]   TARGETING SIGNALS AND SUBUNIT INTERACTIONS IN COATED VESICLE ADAPTER COMPLEXES [J].
PAGE, LJ ;
ROBINSON, MS .
JOURNAL OF CELL BIOLOGY, 1995, 131 (03) :619-630
[18]  
PONNAMBALAM S, 1990, J BIOL CHEM, V265, P4814
[19]  
ROBINSON M S, 1992, Trends in Cell Biology, V2, P293, DOI 10.1016/0962-8924(92)90118-7
[20]   ASSEMBLY AND TARGETING OF ADAPTIN CHIMERAS IN TRANSFECTED CELLS [J].
ROBINSON, MS .
JOURNAL OF CELL BIOLOGY, 1993, 123 (01) :67-77