National Natural Science Foundation of China (NSFC)
31971123
中国
National Natural Science Foundation of China (NSFC)
81920108015
中国
National Natural Science Foundation of China (NSFC)
31930059
中国
引用
ジャーナル: Cell Discov / 年: 2021 タイトル: Mechanism of substrate transport and inhibition of the human LAT1-4F2hc amino acid transporter. 著者: Renhong Yan / Yaning Li / Jennifer Müller / Yuanyuan Zhang / Simon Singer / Lu Xia / Xinyue Zhong / Jürg Gertsch / Karl-Heinz Altmann / Qiang Zhou / 要旨: LAT1 (SLC7A5) is one of the representative light chain proteins of heteromeric amino acid transporters, forming a heterodimer with its heavy chain partner 4F2hc (SLC3A2). LAT1 is overexpressed in ...LAT1 (SLC7A5) is one of the representative light chain proteins of heteromeric amino acid transporters, forming a heterodimer with its heavy chain partner 4F2hc (SLC3A2). LAT1 is overexpressed in many types of tumors and mediates the transfer of drugs and hormones across the blood-brain barrier. Thus, LAT1 is considered as a drug target for cancer treatment and may be exploited for drug delivery into the brain. Here, we synthesized three potent inhibitors of human LAT1, which inhibit transport of leucine with IC values between 100 and 250 nM, and solved the cryo-EM structures of the corresponding LAT1-4F2hc complexes with these inhibitors bound at resolution of up to 2.7 or 2.8 Å. The protein assumes an outward-facing occluded conformation, with the inhibitors bound in the classical substrate binding pocket, but with their tails wedged between the substrate binding site and TM10 of LAT1. We also solved the complex structure of LAT1-4F2hc with 3,5-diiodo-L-tyrosine (Diiodo-Tyr) at 3.4 Å overall resolution, which revealed a different inhibition mechanism and might represent an intermediate conformation between the outward-facing occluded state mentioned above and the outward-open state. To our knowledge, this is the first time that the outward-facing conformation is revealed for the HAT family. Our results unveil more important insights into the working mechanisms of HATs and provide a structural basis for future drug design.
ダウンロード / ファイル: emd_30842.map.gz / 形式: CCP4 / 大きさ: 64 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, focused refined on transmembrane region
ボクセルのサイズ
X=Y=Z: 1.087 Å
密度
表面レベル
登録者による: 0.04 / ムービー #1: 0.04
最小 - 最大
-0.14713943 - 0.2276284
平均 (標準偏差)
-0.00026577065 (±0.0050491695)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
256
256
256
Spacing
256
256
256
セル
A=B=C: 278.272 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
1.087
1.087
1.087
M x/y/z
256
256
256
origin x/y/z
0.000
0.000
0.000
length x/y/z
278.272
278.272
278.272
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
0
0
0
NX/NY/NZ
288
288
288
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
256
256
256
D min/max/mean
-0.147
0.228
-0.000
-
添付データ
-
試料の構成要素
-
全体 : cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, f...
全体
名称: cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, focused refined on transmembrane region
要素
複合体: cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, focused refined on transmembrane region
-
超分子 #1: cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, f...
超分子
名称: cryo EM map of the LAT1-4F2hc bound with 3,5-diiodo-L-tyrosine, focused refined on transmembrane region タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#2
由来(天然)
生物種: Homo sapiens (ヒト)
組換発現
生物種: Homo sapiens (ヒト)
-
実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
緩衝液
pH: 8
凍結
凍結剤: ETHANE
-
電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 50.0 e/Å2