A cell-free biochemical complementation assay reveals complex and redundant cytosolic requirements for LRP endocytosis

被引：3
|
作者：
Miwako, I [1 ]
Schmid, SL [1 ]
机构：
[1] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA
来源：
EXPERIMENTAL CELL RESEARCH | 2006年 / 312卷 / 08期
关键词：
endocytosis; cell-free assay; LDL-receptor related protein; clathrin; dynamin; AP2;
D O I：
10.1016/j.yexcr.2005.12.022
中图分类号：
R73 [肿瘤学];
学科分类号：
100214 ;
摘要：
The low density lipoprotein receptor-related protein (LRP) binds multiple, distinct ligands and participates in constitutive endocytosis and signal transduction. Using an in vitro reconstitution system and a new biochemical complementation assay, we have explored the limiting cytosolic requirements for endocytosis of LRP from isolated plasma membranes. We find that clathrin, AP2 and dynamin do not support efficient LRP uptake and that additional factors present in a 30% ammonium sulfate supernatant fraction of bovine brain cytosol (AS supt) are required. Fractionation of the AS supt revealed that multiple and redundant factors are required to support LRP endocytosis. Among these, we identified Hsc70, synaptojanin1 and CRMP-2 by mass spectrometry. Our data suggest that LRP, which bears several distinct endocytic motifs in its cytoplasmic domain, may use multiple pathways for endocytosis in vitro. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码：1335 / 1344
页数：10
相关论文
共 15 条
  • [1] Northern Lights assay reveals damage to cell-free DNA in sepsis patients
    Helgason, H. H.
    Gudmundsson, B.
    Thormar, H. G.
    Karason, S.
    Sigvaldason, K.
    Jonsson, J. J.
    CLINICA CHIMICA ACTA, 2019, 493 : S580 - S581
  • [2] Biochemical Insights on Degradation of Arabidopsis DELLA Proteins Gained From a Cell-Free Assay System
    Wang, Feng
    Zhu, Danmeng
    Huang, Xi
    Li, Shuang
    Gong, Yinan
    Yao, Qinfang
    Fu, Xiangdong
    Fan, Liu-Min
    Deng, Xing Wang
    PLANT CELL, 2009, 21 (08): : 2378 - 2390
  • [3] Dissecting Functions of the Conserved Oligomeric Golgi Tethering Complex Using a Cell-Free Assay
    Cottam, Nathanael P.
    Wilson, Katherine M.
    Ng, Bobby G.
    Koerner, Christian
    Freeze, Hudson H.
    Ungar, Daniel
    TRAFFIC, 2014, 15 (01) : 12 - 21
  • [4] DEVELOPMENT OF A CELL-FREE ASSAY FROM BOTRYTIS-CINEREA AS A BIOCHEMICAL SCREEN FOR STEROL BIOSYNTHESIS INHIBITORS
    STEHMANN, C
    KAPTEYN, JC
    DEWAARD, MA
    PESTICIDE SCIENCE, 1994, 40 (01): : 1 - 8
  • [5] A cell free protein fragment complementation assay for monitoring the core interaction of the human cytomegalovirus nuclear egress complex
    Schnee, Margit
    Wagner, Felicia M.
    Koszinowski, Ulrich H.
    Ruzsics, Zsolt
    ANTIVIRAL RESEARCH, 2012, 95 (01) : 12 - 18
  • [6] A High-Throughput Genetic Complementation Assay in Yeast Cells Identified Selective Inhibitors of Sphingosine Kinase 1 Not Found Using a Cell-Free Enzyme Assay
    Kashem, Mohammed A.
    Kennedy, Charles A.
    Fogarty, Kylie E.
    Dimock, Janice R.
    Zhang, Yunlong
    Sanville-Ross, Mary L.
    Skow, Donna J.
    Brunette, Steven R.
    Swantek, Jennifer L.
    Hummel, Heidi S.
    Swindle, John
    Nelson, Richard M.
    ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES, 2016, 14 (01) : 39 - 49
  • [7] Cell-based and cell-free firefly luciferase complementation assay to quantify Human Immunodeficiency Virus type 1 Rev-Rev interaction
    Hansen, Tucker
    Baris, Jodie
    Zhao, Min
    Sutton, Richard E.
    VIROLOGY, 2022, 576 : 30 - 41
  • [8] A cell-free mRNA stability assay reveals conservation of the enzymes and mechanisms of mRNA decay between mosquito and mammalian cell lines
    Opyrchal, M
    Anderson, JR
    Sokoloski, KJ
    Wilusz, CJ
    Wilusz, J
    INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 2005, 35 (12) : 1321 - 1334
  • [9] Cell-free Biochemical Fluorometric Enzymatic Assay for High-throughput Measurement of Lipid Peroxidation in High Density Lipoprotein
    Sen Roy, Shubhendu
    Huy Cong Xuan Nguyen
    Angelovich, Thomas A.
    Hearps, Anna C.
    Huynh, Diana
    Jaworowski, Anthony
    Kelesidis, Theodoros
    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2017, (128):
  • [10] THE INHIBITION OF ORGANIC NITRO REDUCTASE BY AUREOMYCIN IN CELL-FREE EXTRACTS .2. COFACTOR REQUIREMENTS FOR THE NITRO REDUCTASE ENZYME COMPLEX
    SAZ, AK
    SLIE, RB
    ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1954, 51 (01) : 5 - 16
12