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Apolipoprotein A-I directly interacts with extracellular domain 1 of human ABCA1

Kawanobe, Takaaki, Shiranaga, Naoko, Kioka, Noriyuki, Kimura, Yasuhisa, Ueda, Kazumitsu·2019·Bioscience, Biotechnology, and Biochemistry

Abstract

ATP-binding cassette transporter A1 (ABCA1) is critical for the generation of nascent high-density lipoprotein (HDL) and plays important roles in cholesterol homeostasis. ABCA1 has two large extracellular domains (ECDs), which may interact directly with apolipoprotein A-I (apoA-I). However, the molecular mechanisms underlying HDL formation and the importance of ABCA1–apoA-I interactions in HDL formation remain unclear. We investigated the ABCA1–apoA-I interaction in photo-activated crosslinking experiments using sulfo-SBED–labeled apoA-I. ApoA-I bound to cells expressing ABCA1, but not to untransfected cells or cells expressing non-functional ABCA1. Binding was inhibited by sulfo-SBED–labeled apoA-I, and crosslinking of sulfo-SBED–labeled apoA-I with ABCA1 was inhibited by non-labeled apoA-I, suggesting that sulfo-SBED–labeled apoA-I specifically binds and crosslinks with functional ABCA1. Proteolytic digestion of crosslinked ABCA1 revealed that apoA-I bound the N-terminal half of ABCA1, and that the first ECD of ABCA1 is an apoA-I binding site. Abbreviations: ABC: ATP-binding cassette; apoA-I: apolipoprotein A-I; ATP: adenosine triphosphate; CHAPS: 3-(3-cholamidepropyl)dimethylammonio-1- propanesulphonate; DTT: dithiothreitol; ECD: extra cellular domain; EDTA: ethylenediaminetetraacetic acid; GFP: green fluorescent protein; HA: hemagglutinin; HDL: high density lipoprotein; HEK: human embryonic kidney; HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; sulfo-SBED: (sulfosuccinimidyl-2-[6-(biotinamido)-2-(p-azidobenzamido)hexanoamido] ethyl-1,3ʹ-dithiopropionate; NHS-ester, N-hydroxysuccinimide-ester

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Crossref · 10.1080/09168451.2018.1547106

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Research works printed but not linked: 26

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Cited by

Metadata

Authors
Kawanobe, T., Shiranaga, N., Kioka, N., Kimura, Y., & Ueda, K.
Affiliations
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan
Venue
Bioscience, Biotechnology, and Biochemistry
Published
2019-03-04
Volume
83
Issue
3
Pages
490-497
ISSN
  • 0916-8451
  • 1347-6947
Cited by
11
References
0
Language
en
Funders
  • AMED-PRIME (JP18gm5910022)
  • Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (15H01638, 15K07390, 18H05269, 18K19176)

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