Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput.
Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput.
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DOI：
10.1128/mbio.03221-21
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发表时间：
2021-12-21
期刊：
影响因子：
6.4
通讯作者：
Bilsland E
中科院分区：
文献类型：
--
作者：
Almeida LD;Silva ASF;Mota DC;Vasconcelos AA;Camargo AP;Pires GS;Furlan M;Freire HMRDC;Klippel AH;Silva SF;Zanelli CF;Carazzolle MF;Oliver SG;Bilsland E
在开发新药时，应该考虑摄取和外排的途径，以便它们能够有效地解决其细胞内靶点。一般来说，药物似乎是通过蛋白质载体进入细胞的，蛋白质载体通常携带营养物质或代谢物。之前开发的一条管道使用携带单基因缺失的酿酒酵母突变株搜索药物转运体，确定了大多数测试化合物的进口路线。然而，由于转运蛋白功能的冗余，我们建议通过在低通量和高通量筛选中利用双突变菌株来改进这一方法。我们构建了一个包含122个非必需质膜转运蛋白编码基因双缺失的14,000多株菌株的文库，并进行了低通量和高通量筛选，确定了23种化合物可能的药物进口途径。此外，高通量分析能够确定21种化合物的推测外排途径。集中在唑类抗真菌药物上，我们能够确定肌醇转运体Itr1p参与这些分子的摄取，并确认Pdr5p在它们的输出中的作用。
The routes of uptake and efflux should be considered when developing new drugs so that they can effectively address their intracellular targets. As a general rule, drugs appear to enter cells via protein carriers that normally carry nutrients or metabolites. A previously developed pipeline that searched for drug transporters using Saccharomyces cerevisiae mutants carrying single-gene deletions identified import routes for most compounds tested. However, due to the redundancy of transporter functions, we propose that this methodology can be improved by utilizing double mutant strains in both low- and high-throughput screens. We constructed a library of over 14,000 strains harboring double deletions of genes encoding 122 nonessential plasma membrane transporters and performed low- and high-throughput screens identifying possible drug import routes for 23 compounds. In addition, the high-throughput assay enabled the identification of putative efflux routes for 21 compounds. Focusing on azole antifungals, we were able to identify the involvement of the myo-inositol transporter, Itr1p, in the uptake of these molecules and to confirm the role of Pdr5p in their export.
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影响因子：
48
作者：
Callahan BJ;McMurdie PJ;Rosen MJ;Han AW;Johnson AJ;Holmes SP
通讯作者：
Holmes SP
影响因子：
56.9
作者：
Hillenmeyer, Maureen E.;Fung, Eula;Giaever, Guri
通讯作者：
Giaever, Guri
DOI：
10.1111/j.1432-1033.1988.tb13806.x
发表时间：
1988-01-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
JUND, R;WEBER, E;CHEVALLIER, MR
通讯作者：
CHEVALLIER, MR
影响因子：
6.7
作者：
Hoepfner, Dominic;Helliwell, Stephen B.;Movva, N. Rao
通讯作者：
Movva, N. Rao
影响因子：
5.2
作者：
Esquivel BD;White TC
通讯作者：
White TC