Redox-dependent internalization of the purinergic P2Y6 receptor limits colitis progression
Redox-dependent internalization of the purinergic P2Y6 receptor limits colitis progression
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DOI：
10.1126/scisignal.abj0644
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发表时间：
2022-01-11
影响因子：
7.3
通讯作者：
Nishida, Motohiro
中科院分区：
文献类型：
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作者：
Nishiyama, Kazuhiro;Nishimura, Akiyuki;Nishida, Motohiro
在配体刺激后，许多G蛋白偶联受体（GPCR）经历β-抑制蛋白依赖性脱敏，在此期间它们被内化并降解或再循环至质膜。一些GPCR不受这种类型的脱敏作用的影响，因为它们缺乏与β-抑制蛋白相互作用所需的残基。我们确定了一种氧化还原依赖性替代内化（REDAI）机制，该机制促进嘌呤能P2 Y（6）受体（P2 Y（6）R）的内化和降解。含有亲电子异硫氰酸酯基团的合成和天然化合物共价修饰了Cys（220）处的P2 Y（6）R，这促进了各种小鼠和人类培养细胞系中Lys（137）的泛素化以及受体内化和降解。内源性亲电体也促进配体依赖性P2 Y（6）R的内化和降解。P2 Y（6）R在炎症细胞中含量丰富，促进结肠炎的发病机制。P2 Y（6）R的缺陷保护小鼠免受实验诱导的结肠炎，并且表达P2 Y（6）R形式的小鼠对结肠炎的诱导更敏感，其中Cys（220）突变为不可修饰的丝氨酸。其他几种GPCR，包括A（2B）AR，在适当位置含有半胱氨酸和赖氨酸残基以介导REDAI，异硫氰酸酯刺激A（2B）AR和插入适当残基的P2 Y（2）R形式的内化。因此，内源性和外源性亲电体可以通过P2 Y（6）R的半胱氨酸修饰来限制结肠炎进展，并且还可以介导其他GPCR的内化。
After ligand stimulation, many G protein-coupled receptors (GPCRs) undergo beta-arrestin-dependent desensitization, during which they are internalized and either degraded or recycled to the plasma membrane. Some GPCRs are not subject to this type of desensitization because they lack the residues required to interact with beta-arrestins. We identified a mechanism of redox-dependent alternative internalization (REDAI) that promotes the internalization and degradation of the purinergic P2Y(6) receptor (P2Y(6)R). Synthetic and natural compounds containing electrophilic isothiocyanate groups covalently modified P2Y(6)R at Cys(220), which promoted the ubiquitylation of Lys(137) and receptor internalization and degradation in various mouse and human cultured cell lines. Endogenous electrophiles also promoted ligand-dependent P2Y(6)R internalization and degradation. P2Y(6)R is highly abundant in inflammatory cells and promotes the pathogenesis of colitis. Deficiency in P2Y(6)R protected mice against experimentally induced colitis, and mice expressing a form of P2Y(6)R in which Cys(220) was mutated to nonmodifiable serine were more sensitive to the induction of colitis. Several other GPCRs, including A(2B)AR, contain cysteine and lysine residues at the appropriate positions to mediate REDAI, and isothiocyanate stimulated the internalization of A(2B)AR and of a form of P2Y(2)R with insertions of the appropriate residues. Thus, endogenous and exogenous electrophiles may limit colitis progression through cysteine modification of P2Y(6)R and may also mediate internalization of other GPCRs.