Identification of secreted proteins by comparison of protein abundance in conditioned media and cell lysates.
Identification of secreted proteins by comparison of protein abundance in conditioned media and cell lysates.
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DOI：
10.1016/j.ab.2022.114846
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发表时间：
2022-10-15
影响因子：
2.9
通讯作者：
中科院分区：
文献类型：
--
作者：
细胞培养物中分泌蛋白质的全谱分析由于细胞内蛋白质从受损细胞的泄漏而变得复杂。为了解决这个问题，我们比较了细胞裂解物和条件培养基之间的单个蛋白质的丰度，推断分泌的蛋白质在条件培养基中应该相对更丰富。随着条件培养基与细胞裂解物比率的增加，携带信号肽的蛋白质的显著富集，以及布雷菲德菌素A处理后该信号的丧失，证实了该方法的灵敏度和特异性。在布雷菲德菌素A存在下，表现出条件培养基与细胞裂解物比率增加的蛋白质亚组鉴定了通过不依赖于ER至高尔基体运输的途径进行非常规分泌的候选物。
Analysis of the full spectrum of secreted proteins in cell culture is complicated by leakage of intracellular proteins from damaged cells. To address this issue, we compared the abundance of individual proteins between the cell lysate and the conditioned medium, reasoning that secreted proteins should be relatively more abundant in the conditioned medium. Marked enrichment for signal-peptide-bearing proteins with increasing conditioned media to cell lysate ratio, as well loss of this signal following brefeldin A treatment, confirmed the sensitivity and specificity of this approach. The subset of proteins demonstrating increased conditioned media to cell lysate ratio in the presence of Brefeldin A identified candidates for unconventional secretion via a pathway independent of ER to Golgi trafficking.
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影响因子：
4.4
作者：
Grube, Leonie;Dellen, Rafael;Poschmann, Gereon
通讯作者：
Poschmann, Gereon
影响因子：
4.4
作者：
Loei, Hendrick;Tan, Hwee Tong;Chung, Maxey C. M.
通讯作者：
Chung, Maxey C. M.
影响因子：
64.8
作者：
Popow J;Jurkin J;Schleiffer A;Martinez J
通讯作者：
Martinez J
影响因子：
7.2
作者：
Grieve, Adam G.;Rabouille, Catherine
通讯作者：
Rabouille, Catherine
DOI：
10.1007/978-1-4939-9133-4_19
发表时间：
2019-01-01
期刊：
EXTRACELLULAR MATRIX
影响因子：
--
作者：
Purushothaman, Anurag
通讯作者：
Purushothaman, Anurag