当前位置: X-MOL 学术Bioconjugate Chem. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Isotope-Coded Maleimide Affinity Tags for Proteomics Applications
Bioconjugate Chemistry ( IF 4.7 ) Pub Date : 2021-06-23 , DOI: 10.1021/acs.bioconjchem.1c00206
Adam P Wdowiak , Marisa N Duong 1 , Rohan D Joyce , Amber E Boyatzis , Mark C Walkey , Gareth L Nealon , Peter G Arthur , Matthew J Piggott
Affiliation  

Isotope-coded affinity tags (ICATs) are valuable tools for mass spectrometry-based quantitative proteomics, in particular, for comparison of protein (cysteine-residue) thiol oxidation state in normal, stressed, and diseased tissue. However, the iodoacetamido electrophile used in most commercial ICATs suffers from poor thiol-selectivity and modest rates of adduct formation, which can lead to spurious results. Hence, we designed and synthesized three ICATs containing thiol-selective N-alkylmaleimide electrophiles (isotope-coded maleimide affinity tags = ICMATs) and assessed these as mass spectrometry probes for ratiometric analysis of lysozyme and muscle proteomes. Two ICMAT pairs containing butylene/D8-butylene linkers were effective MS probes, but not ideal for typical proteomics workflows, because peptides bearing these tags frequently did not coelute with HPLC. A switch to a phenylene/13C6-phenylene linker solved this issue without compromising the efficiency of adduct formation.

中文翻译:

用于蛋白质组学应用的同位素编码马来酰亚胺亲和标签

同位素编码的亲和标签 (ICAT) 是基于质谱的定量蛋白质组学的宝贵工具,特别是用于比较正常、应激和患病组织中的蛋白质(半胱氨酸残基)硫醇氧化状态。然而,在大多数商业 ICAT 中使用的碘乙酰胺亲电试剂具有较差的硫醇选择性和适度的加合物形成率,这可能导致虚假结果。因此,我们设计并合成了三种含有硫醇选择性N-烷基马来酰亚胺亲电试剂(同位素编码的马来酰亚胺亲和标签 = ICMAT)的 ICAT,并将它们评估为用于溶菌酶和肌肉蛋白质组比例分析的质谱探针。两个包含丁烯/D 8 的ICMAT 对-丁烯接头是有效的 MS 探针,但对于典型的蛋白质组学工作流程而言并不理想,因为带有这些标签的肽段通常不会与 HPLC 共流出。转换为亚苯基/ 13 C 6 -亚苯基接头解决了这个问题,同时不影响加合物形成的效率。
更新日期:2021-08-19
down
wechat
bug