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Regulation and function of the protein inhibitor of nitric oxide synthase (PIN)/dynein light chain 8 (LC8) in a human mast cell line.

机译:一氧化氮合酶(PIN)/动力蛋白轻链8(LC8)在人类肥大细胞系中的调节和功能。

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摘要

The protein inhibitor of nitric oxide synthase (PIN) was independently identified as an inhibitor of nitric oxide (NO) produced by neuronal nitric oxide synthase (nNOS), and as a member of the cellular dynein light chain family, dynein light chain 8 (LC8), responsible for intracellular protein trafficking. Mast cells (MC) are involved in several homeostatic and pathological processes and can be regulated by NO. This study describes the expression of PIN/LC8 in the human MC line HMC-1. We also studied if PIN/LC8 binds nNOS, and what role this might have in leukotriene (LT) production. We found that PIN/LC8 mRNA and protein was expressed in HMC-1. Using a GST-PIN construct, we showed PIN binds to nNOS, but not endothelial (e)NOS in HMC-1; in our studies HMC-1 did not express inducible (i)NOS. Intracellular delivery of anti-PIN/LC8 antibody enhanced ionophore (A23187)-induced LT production through an unknown mechanism. Thus we established for the first time expression of PIN/LC8 in human MC, its ability to bind nNOS, and the effect that blocking it has on LT production in a human MC lines.
机译:一氧化氮合酶(PIN)的蛋白抑制剂被独立鉴定为由神经元一氧化氮合酶(nNOS)产生的一氧化氮(NO)抑制剂,并且是细胞达因轻链8轻链家族(LC8)的成员),负责细胞内蛋白质的运输。肥大细胞(MC)参与多个体内平衡和病理过程,并可由NO调节。这项研究描述了PIN / LC8在人MC系HMC-1中的表达。我们还研究了PIN / LC8是否结合nNOS,以及这可能在白三烯(LT)生产中发挥什么作用。我们发现PIN / LC8 mRNA和蛋白在HMC-1中表达。使用GST-PIN构建体,我们显示PIN与HMC-1中的nNOS结合,但不与内皮(e)NOS结合。在我们的研究中,HMC-1不表达诱导型(i)NOS。细胞内递送抗PIN / LC8抗体通过未知机制增强了离子载体(A23187)诱导的LT产生。因此,我们首次确定了PIN / LC8在人MC中的表达,其结合nNOS的能力以及阻断它对人MC系中LT产生的影响。

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