Dynamic and redundant regulation of LRRK2 and LRRK1 expression.
Dynamic and redundant regulation of LRRK2 and LRRK1 expression.
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DOI：
10.1186/1471-2202-8-102
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发表时间：
2007-11-28
期刊：
影响因子：
2.4
通讯作者：
West AB
中科院分区：
文献类型：
--
作者：
Biskup S;Moore DJ;Rea A;Lorenz-Deperieux B;Coombes CE;Dawson VL;Dawson TM;West AB
富含亮氨酸的重复序列激酶2（LRRK 2）基因内的突变占常染色体显性遗传和一些迟发性散发性帕金森病的很大比例。阐明LRRK 2蛋白在健康和疾病中的功能为破译神经变性中重要的分子途径提供了机会。在哺乳动物中，LRRK 1和LRRK 2蛋白包含编码控制内在激酶活性的GT3结构域的独特家族。鼠LRRK蛋白的表达谱尚未完全描述，并且未充分表征的抗体在文献中产生了相互矛盾的结果。在此，我们使用小鼠LRRK 2和人LRRK 2表达载体、野生型和LRRK 2缺失小鼠脑裂解物和人脑裂解物全面评估了21种市售的LRRK 2蛋白抗体。11种抗体检测过表达的人LRRK 2，而4种抗体检测内源性人LRRK 2。相反，两种抗体识别过表达的小鼠LRRK 2，一种抗体检测内源性小鼠LRRK 2。LRRK 2蛋白存在于可溶性和去污剂可溶性蛋白级分中。LRRK 2和相关的LRRK 1基因编码低水平的表达mRNA种类，对应于发育期间和成年期的低水平蛋白质，具有很大程度上冗余的表达谱。尽管先前公布的结果，市售抗体通常不能识别内源性小鼠LRRK 2蛋白;然而，几种抗体保留了检测过表达的小鼠LRRK 2蛋白的能力。超过一半的市售抗体检测过表达的人LRRK 2蛋白，一些具有足够的特异性来检测人脑中的内源性LRRK 2。哺乳动物LRRK蛋白在几种组织中受到发育调控，协调表达表明LRRK 1和LRRK 2之间的功能可能存在冗余。
Mutations within the leucine-rich repeat kinase 2 (LRRK2) gene account for a significant proportion of autosomal-dominant and some late-onset sporadic Parkinson's disease. Elucidation of LRRK2 protein function in health and disease provides an opportunity for deciphering molecular pathways important in neurodegeneration. In mammals, LRRK1 and LRRK2 protein comprise a unique family encoding a GTPase domain that controls intrinsic kinase activity. The expression profiles of the murine LRRK proteins have not been fully described and insufficiently characterized antibodies have produced conflicting results in the literature. Herein, we comprehensively evaluate twenty-one commercially available antibodies to the LRRK2 protein using mouse LRRK2 and human LRRK2 expression vectors, wild-type and LRRK2-null mouse brain lysates and human brain lysates. Eleven antibodies detect over-expressed human LRRK2 while four antibodies detect endogenous human LRRK2. In contrast, two antibodies recognize over-expressed mouse LRRK2 and one antibody detected endogenous mouse LRRK2. LRRK2 protein resides in both soluble and detergent soluble protein fractions. LRRK2 and the related LRRK1 genes encode low levels of expressed mRNA species corresponding to low levels of protein both during development and in adulthood with largely redundant expression profiles. Despite previously published results, commercially available antibodies generally fail to recognize endogenous mouse LRRK2 protein; however, several antibodies retain the ability to detect over-expressed mouse LRRK2 protein. Over half of the commercially available antibodies tested detect over-expressed human LRRK2 protein and some have sufficient specificity to detect endogenous LRRK2 in human brain. The mammalian LRRK proteins are developmentally regulated in several tissues and coordinated expression suggest possible redundancy in the function between LRRK1 and LRRK2.
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影响因子：
11.2
作者：
Biskup, Saskia;Moore, Darren J.;Dawson, Valina L.
通讯作者：
Dawson, Valina L.
影响因子：
3.3
作者：
Melrose, H;Lincoln, S;Farrer, M
通讯作者：
Farrer, M
影响因子：
10.7
作者：
Marin, Ignacio
通讯作者：
Marin, Ignacio
影响因子：
6.1
作者：
Greggio, Elisa;Jain, Shushant;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
11.2
作者：
Giasson, BI;Covy, JP;Van Deerlin, VM
通讯作者：
Van Deerlin, VM