Dominici S, Fiori V, Magnani M, Schena E, Capanni C, Camozzi D, D'Apice M R, Le Dour C, Auclair M, Caron M, Novelli G, Vigouroux C, Maraldi N M, Lattanzi G
Institute of Biochemistry, University of Urbino, Urbino, Italy.
Eur J Histochem. 2009 Jan-Mar;53(1):43-52. doi: 10.4081/ejh.2009.43.
Lamin A is a component of the nuclear lamina mutated in a group of human inherited disorders known as laminopathies. Among laminopathies, progeroid syndromes and lipodystrophies feature accumulation of prelamin A, the precursor protein which, in normal cells, undergoes a multi-step processing to yield mature lamin A. It is of utmost importance to characterize the prelamin A form accumulated in each laminopathy, since existing evidence shows that drugs acting on protein processing can improve some pathological aspects.We report that two antibodies raised against differently modified prelamin A peptides show a clear specificity to full-length prelamin A or carboxymethylated farnesylated prelamin A, respectively. Using these antibodies, we demonstrated that inhibition of the prelamin A endoprotease ZMPSTE24 mostly elicits accumulation of full-length prelamin A in its farnesylated form, while loss of the prelamin A cleavage site causes accumulation of carboxymethylated prelamin A in progeria cells. These results suggest a major role of ZMPSTE24 in the first prelamin A cleavage step.
核纤层蛋白A是核纤层的一个组成部分,在一组称为核纤层蛋白病的人类遗传性疾病中发生突变。在核纤层蛋白病中,早衰综合征和脂肪营养不良的特征是前体核纤层蛋白A的积累,这种前体蛋白在正常细胞中会经过多步加工以产生成熟的核纤层蛋白A。表征每种核纤层蛋白病中积累的前体核纤层蛋白A形式至关重要,因为现有证据表明作用于蛋白质加工的药物可以改善某些病理状况。我们报告,针对不同修饰的前体核纤层蛋白A肽产生的两种抗体分别对全长前体核纤层蛋白A或羧甲基化法尼基化前体核纤层蛋白A具有明显的特异性。使用这些抗体,我们证明前体核纤层蛋白A内切蛋白酶ZMPSTE24的抑制主要引发全长法尼基化形式的前体核纤层蛋白A的积累,而前体核纤层蛋白A切割位点的缺失导致早衰细胞中羧甲基化前体核纤层蛋白A的积累。这些结果表明ZMPSTE24在前体核纤层蛋白A的第一步切割中起主要作用。