XPA is susceptible to proteolytic cleavage by cathepsin L during lysis of quiescent cells.
XPA is susceptible to proteolytic cleavage by cathepsin L during lysis of quiescent cells.
复制标题
DOI：
10.1016/j.dnarep.2021.103260
复制
发表时间：
2022-01
期刊：
影响因子：
3.8
通讯作者：
Kemp MG
中科院分区：
文献类型：
--
作者：
Khan S;Cvammen W;Anabtawi N;Choi JH;Kemp MG
着色性干皮病A组（XPA）蛋白作为核苷酸切除修复（NER）机制的一个组成部分，在从DNA中去除UV光产物和其他大块病变中起着至关重要的作用。使用从人细胞和人皮肤表皮的融合培养物制备的细胞裂解物，我们在免疫印迹上观察到比天然全长XPA蛋白小约3- 4kDa的额外XPA抗体反应性条带。生物化学研究表明，这种较小分子量的XPA物质是由于蛋白质C末端的蛋白水解，这对XPA与NER蛋白TFIIH相互作用的能力产生了负面影响。进一步的工作鉴定了内肽酶组织蛋白酶L，其在静止细胞中以较高水平表达，作为负责在细胞裂解期间裂解XPA的蛋白酶。这些结果表明，补充具有组织蛋白酶L抑制剂的裂解缓冲液对于防止融合细胞裂解期间XPA的裂解是重要的。
The xeroderma pigmentosum group A (XPA) protein plays an essential role in the removal of UV photoproducts and other bulky lesions from DNA as a component of the nucleotide excision repair (NER) machinery. Using cell lysates prepared from confluent cultures of human cells and from human skin epidermis, we observed an additional XPA antibody-reactive band on immunoblots that was approximately 3–4 kDa smaller than the native, full-length XPA protein. Biochemical studies revealed this smaller molecular weight XPA species to be due to proteolysis at the C-terminus of the protein, which negatively impacted the ability of XPA to interact with the NER protein TFIIH. Further work identified the endopeptidase cathepsin L, which is expressed at higher levels in quiescent cells, as the protease responsible for cleaving XPA during cell lysis. These results suggest that supplementation of lysis buffers with inhibitors of cathepsin L is important to prevent cleavage of XPA during lysis of confluent cells.
登录
查看更多内容
影响因子：
11.4
作者：
Gonzalez-Suarez, Ignacio;Redwood, Abena B.;Gonzalo, Susana
通讯作者：
Gonzalo, Susana
影响因子：
64.8
作者：
LOCKWOOD, TD;SHIER, WT
通讯作者：
SHIER, WT
影响因子：
3.8
作者：
Manandhar, Mandira;Lowery, Megan G.;Wood, Richard D.
通讯作者：
Wood, Richard D.
影响因子：
11.2
作者：
Adair, Jennifer E.;Maloney, Scott C.;Reeves, Raymond
通讯作者：
Reeves, Raymond
影响因子：
8
作者：
Iakoucheva, LM;Kimzey, AL;Ackerman, EJ
通讯作者：
Ackerman, EJ