Senescence marker protein 30 (SMP30) expression in eukaryotic cells: existence of multiple species and membrane localization.
Senescence marker protein 30 (SMP30) expression in eukaryotic cells: existence of multiple species and membrane localization.
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DOI：
10.1371/journal.pone.0016545
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发表时间：
2011-02-09
期刊：
影响因子：
3.7
通讯作者：
Chilukuri N
中科院分区：
文献类型：
--
作者：
Arun P;Aleti V;Parikh K;Manne V;Chilukuri N
衰老标志蛋白（SMP 30），又称regucalcin，是一种分子量为34 kDa的细胞质衰老标志蛋白，在细胞内Ca 2+稳态、抗坏血酸生物合成、氧化应激和化学战神经毒剂的解毒中起重要作用。为了研究SMP 30对神经毒剂的解毒活性，我们制备了一种表达人SMP 30的重组腺病毒，该重组腺病毒为带有血凝素标签的融合蛋白（Ad-SMP 30-HA）。Ad-SMP 30-HA以剂量和时间依赖的方式转导HEK-293 A和C3 A肝细胞中SMP 30-HA和另外两种分子大小分别为约28 kDa和24 kDa的SMP 30的表达。在小鼠中静脉内施用Ad-SMP 30-HA导致所有三种形式的SMP 30在肝脏和膈肌中表达。LC-MS/MS结果证实，较低分子量的28 kDa和24 kDa蛋白质与34 kDa SMP 30相关。在正常大鼠肝脏和注射Ad-SMP 30-HA的小鼠中也检测到28 kDa和24 kDa的SMP 30形式，表明SMP 30在生理条件下确实以多种形式存在。在两种细胞系中的时程实验表明，28 kDa和24 kDa SMP 30形式可能由34 kDa SMP 30产生。有趣的是，28 kDa和24 kDa的SMP 30形式最初出现在细胞质中，并转移到颗粒部分。使用蛋白水解途径的小分子抑制剂的研究揭示了β和γ-分泌酶的潜在参与，但不包括钙蛋白酶、溶酶体蛋白酶、蛋白酶体和半胱天冬酶。这是第一份报告描述了多种形式的SMP 30的存在，它们的优先分布膜和它们的产生可能通过分泌酶介导的蛋白水解。
Senescence marker protein (SMP30), also known as regucalcin, is a 34 kDa cytosolic marker protein of aging which plays an important role in intracellular Ca2+ homeostasis, ascorbic acid biosynthesis, oxidative stress, and detoxification of chemical warfare nerve agents. In our goal to investigate the activity of SMP30 for the detoxification of nerve agents, we have produced a recombinant adenovirus expressing human SMP30 as a fusion protein with a hemaglutinin tag (Ad-SMP30-HA). Ad-SMP30-HA transduced the expression of SMP30-HA and two additional forms of SMP30 with molecular sizes ∼28 kDa and 24 kDa in HEK-293A and C3A liver cells in a dose and time-dependent manner. Intravenous administration of Ad-SMP30-HA in mice results in the expression of all the three forms of SMP30 in the liver and diaphragm. LC-MS/MS results confirmed that the lower molecular weight 28 kDa and 24 kDa proteins are related to the 34 kDa SMP30. The 28 kDa and 24 kDa SMP30 forms were also detected in normal rat liver and mice injected with Ad-SMP30-HA suggesting that SMP30 does exist in multiple forms under physiological conditions. Time course experiments in both cell lines suggest that the 28 kDa and 24 kDa SMP30 forms are likely generated from the 34 kDa SMP30. Interestingly, the 28 kDa and 24 kDa SMP30 forms appeared initially in the cytosol and shifted to the particulate fraction. Studies using small molecule inhibitors of proteolytic pathways revealed the potential involvement of β and γ-secretases but not calpains, lysosomal proteases, proteasome and caspases. This is the first report describing the existence of multiple forms of SMP30, their preferential distribution to membranes and their generation through proteolysis possibly mediated by secretase enzymes.
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DOI：
10.1196/annals.1297.062
发表时间：
2004-01-01
期刊：
STRATEGIES FOR ENGINEERED NEGLIGIBLE SENESCENCE: WHY GENUINE CONTROL OF AGING MAY BE FORESEEABLE
影响因子：
--
作者：
Feng, DY;Kondo, Y;Maruyama, N
通讯作者：
Maruyama, N
影响因子：
14.9
作者：
Leparc, German Gaston;Mitra, Robi David
通讯作者：
Mitra, Robi David
影响因子：
4
作者：
Nakagawa, Taeko;Yamaguchi, Masayoshi
通讯作者：
Yamaguchi, Masayoshi
DOI：
10.1006/bbrc.1998.9841
发表时间：
1999-01-08
影响因子：
3.1
作者：
Fujita, T
通讯作者：
Fujita, T
影响因子：
56.9
作者：
ROGERS, S;WELLS, R;RECHSTEINER, M
通讯作者：
RECHSTEINER, M