PMID- 34942188
OWN - NLM
STAT- MEDLINE
DCOM- 20220214
LR  - 20220214
IS  - 1090-2422 (Electronic)
IS  - 0014-4827 (Linking)
VI  - 411
IP  - 1
DP  - 2022 Feb 1
TI  - The two-domain architecture of LAMP2A regulates its interaction with Hsc70.
PG  - 112986
LID - S0014-4827(21)00542-5 [pii]
LID - 10.1016/j.yexcr.2021.112986 [doi]
AB  - Chaperone-mediated autophagy (CMA) is a unique proteolytic pathway, in which 
      cytoplasmic proteins recognized by heat shock cognate protein 70 (Hsc70/HSPA8) 
      are transported into lysosomes for degradation. The substrate/chaperone complex 
      binds to the cytosolic tail of the lysosomal-associated membrane protein type 2A 
      (LAMP2A), but whether the interaction between Hsc70 and LAMP2A is direct or 
      mediated by other molecules has remained to be elucidated. The structure of 
      LAMP2A comprises a large lumenal domain composed of two domains, both with the 
      beta-prism fold, a transmembrane domain and a short cytoplasmic tail. We previously 
      reported the structural basis for the homophilic interaction of the lumenal 
      domains of LAMP2A, using site-specific photo-crosslinking and/or steric hindrance 
      within cells. In the present study, we introduced a photo-crosslinker into the 
      cytoplasmic tail of LAMP2A and successfully detected its crosslinking with Hsc70, 
      revealing this direct interaction for the first time. Furthermore, we 
      demonstrated that the truncation of the membrane-distal domain within the lumenal 
      domain of LAMP2A reduced the amount of Hsc70 that coimmunoprecipitated with 
      LAMP2A. Our present results suggested that the two-domain architecture of the 
      lumenal domains of LAMP2A underlies the interaction with Hsc70 at the cytoplasmic 
      surface of the lysosome.
CI  - Copyright (c) 2021. Published by Elsevier Inc.
FAU - Ikami, Yuta
AU  - Ikami Y
AD  - Department of Oral and Maxillofacial Surgery, Graduate School of Medical and 
      Dental Sciences, Tokyo Medical and Dental University (TMDU), Yushima 1-5-45, 
      Bunkyo-ku, Tokyo, 113-8549, Japan; Department of Biochemistry, Graduate School of 
      Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Yushima 
      1-5-45, Bunkyo-ku, Tokyo, 113-8549, Japan.
FAU - Terasawa, Kazue
AU  - Terasawa K
AD  - Department of Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo 
      Medical and Dental University (TMDU), Yushima 1-5-45, Bunkyo-ku, Tokyo, 113-8549, 
      Japan.
FAU - Sakamoto, Kensaku
AU  - Sakamoto K
AD  - RIKEN Center for Biosystems Dynamics Research (BDR), 1-7-22 Suehiro-cho, 
      Tsurumi-ku, Yokohama, 230-0045, Japan.
FAU - Ohtake, Kazumasa
AU  - Ohtake K
AD  - RIKEN Center for Biosystems Dynamics Research (BDR), 1-7-22 Suehiro-cho, 
      Tsurumi-ku, Yokohama, 230-0045, Japan.
FAU - Harada, Hiroyuki
AU  - Harada H
AD  - Department of Oral and Maxillofacial Surgery, Graduate School of Medical and 
      Dental Sciences, Tokyo Medical and Dental University (TMDU), Yushima 1-5-45, 
      Bunkyo-ku, Tokyo, 113-8549, Japan.
FAU - Watabe, Tetsuro
AU  - Watabe T
AD  - Department of Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo 
      Medical and Dental University (TMDU), Yushima 1-5-45, Bunkyo-ku, Tokyo, 113-8549, 
      Japan.
FAU - Yokoyama, Shigeyuki
AU  - Yokoyama S
AD  - RIKEN Cluster for Science, Technology and Innovation Hub, 1-7-22 Suehiro-cho, 
      Tsurumi-ku, Yokohama, 230-0045, Japan.
FAU - Hara-Yokoyama, Miki
AU  - Hara-Yokoyama M
AD  - Department of Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo 
      Medical and Dental University (TMDU), Yushima 1-5-45, Bunkyo-ku, Tokyo, 113-8549, 
      Japan. Electronic address: m.yokoyama.bch@tmd.ac.jp.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20211221
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (HSC70 Heat-Shock Proteins)
RN  - 0 (HSPA8 protein, human)
RN  - 0 (LAMP2 protein, human)
RN  - 0 (Lysosomal-Associated Membrane Protein 2)
SB  - IM
MH  - Cross-Linking Reagents/*metabolism
MH  - Cytoplasm/*metabolism
MH  - HSC70 Heat-Shock Proteins/chemistry/*metabolism
MH  - Humans
MH  - Lysosomal-Associated Membrane Protein 2/chemistry/*metabolism
MH  - Lysosomes/*metabolism
MH  - *Protein Interaction Domains and Motifs
OTO - NOTNLM
OT  - Chaperone-mediated autophagy
OT  - Expanded genetic codes
OT  - Lysosomes
OT  - Photo-crosslinking
OT  - Protein assembly
EDAT- 2021/12/24 06:00
MHDA- 2022/02/15 06:00
CRDT- 2021/12/23 20:12
PHST- 2021/08/22 00:00 [received]
PHST- 2021/12/11 00:00 [revised]
PHST- 2021/12/19 00:00 [accepted]
PHST- 2021/12/24 06:00 [pubmed]
PHST- 2022/02/15 06:00 [medline]
PHST- 2021/12/23 20:12 [entrez]
AID - S0014-4827(21)00542-5 [pii]
AID - 10.1016/j.yexcr.2021.112986 [doi]
PST - ppublish
SO  - Exp Cell Res. 2022 Feb 1;411(1):112986. doi: 10.1016/j.yexcr.2021.112986. Epub 
      2021 Dec 21.