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Acid sphingomyelinase regulates the localization and trafficking of palmitoylated proteins

机译:酸性鞘磷脂酶调节棕榈酰化蛋白的定位和运输

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In human, loss of acid sphingomyelinase (ASM/SMPD1) causes Niemann–Pick disease, type A. ASM hydrolyzes sphingomyelins to produce ceramides but protein targets of ASM remain largely unclear. Our mass spectrometry-based proteomic analyses have identified 100 proteins associated with the ASM-dependent, detergent-resistant membrane microdomains (lipid rafts), with 60% of these proteins being palmitoylated, including SNAP23, Src-family kinases Yes and Lyn, and Ras and Rab family small GTPases. Inactivation of ASM abolished the presence of these proteins in the plasma membrane, with many of them trapped in the Golgi. While palmitoylation inhibitors and palmitoylation mutants phenocopied the effects of ASM inactivation, we demonstrated that ASM is required for the transport of palmitoylated proteins, such as SNAP23 and Lyn, from the Golgi to the plasma membrane without affecting palmitoylation directly. Importantly, ASM delivered extracellularly can regulate the trafficking of SNAP23 from the Golgi to the plasma membrane. Our studies suggest that ASM, acting at the plasma membrane to produce ceramides, regulates the localization and trafficking of the palmitoylated proteins.
机译:在人类中,酸性鞘磷脂酶(ASM / SMPD1)的丧失会导致Niemann–Pick病(A型)。ASM水解鞘磷脂产生神经酰胺,但ASM的蛋白质靶标仍然不清楚。我们基于质谱的蛋白质组学分析已鉴定出> 100种与ASM依赖的,耐去污剂的膜微结构域(脂质筏)相关的蛋白质,其中> 60%的蛋白质被棕榈酸酯化,包括SNAP23,Src家族激酶Yes和Lyn,以及Ras和Rab家族的小型GTPases。 ASM的失活消除了质膜中这些蛋白质的存在,其中许多蛋白质被困在高尔基体中。虽然棕榈酰化抑制剂和棕榈酰化突变体表型化了ASM失活的作用,但我们证明ASM是棕榈酰化蛋白(例如SNAP23和Lyn)从高尔基体到质膜的运输所必需的,而又不直接影响棕榈酰化。重要的是,细胞外递送的ASM可以调节SNAP23从高尔基体到质膜的运输。我们的研究表明,ASM在质膜上产生神经酰胺,可调节棕榈酰化蛋白的定位和运输。

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