Digestion of native proteins for proteomics using a thermocycler
Digestion of native proteins for proteomics using a thermocycler
复制标题
DOI：
10.1021/ac702527b
复制
发表时间：
2008-08-01
影响因子：
7.4
通讯作者：
Pangburn, Michael K.
中科院分区：
文献类型：
--
作者：
Turapov, Obolbek A.;Mukamolova, Galina V.;Pangburn, Michael K.
Efficient protein digestion is a critical step for successful mass spectrometry analysis. Here we describe simultaneous tryptic digestion and gradual unfolding of native proteins by application of a temperature gradient using a single cycle of 5 min or less in a PCR thermocycler. Chemicals typically used for chromatographic techniques did not affect the digestion efficiency. Tryptic digestion was performed in a small volume (3 mu L) with 1.5 mu g of trypsin without denaturing agents. This rapid procedure yielded more peptides than conventional methods utilizing chemical denaturation for 18 proteins out of 20. Samples were directly spotted on the MALDI-TOF target plate, without additional purification, thus reducing losses on reversed-phase resins.
