1] Centre for Blood Research, University of British Columbia, Vancouver, British Columbia, Canada. [2] Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, British Columbia, Canada. [3] Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada. [4] Zentralinstitut für Engineering, Elektronik und Analytik, ZEA-3: Analytik, Forschungszentrum Jülich, Jülich, Germany.
1] Centre for Blood Research, University of British Columbia, Vancouver, British Columbia, Canada. [2] Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, British Columbia, Canada. [3] Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
Nat Methods. 2015 Jan;12(1):55-8. doi: 10.1038/nmeth.3177. Epub 2014 Nov 24.
To improve proteome coverage and protein C-terminal identification, we characterized the Methanosarcina acetivorans thermophilic proteinase LysargiNase, which cleaves before lysine and arginine up to 55 °C. Unlike trypsin, LysargiNase-generated peptides had N-terminal lysine or arginine residues and fragmented with b ion-dominated spectra. This improved protein C terminal-peptide identification and several arginine-rich phosphosite assignments. Notably, cleavage also occurred at methylated or dimethylated lysine and arginine, facilitating detection of these epigenetic modifications.
为了提高蛋白质组覆盖度和蛋白质 C 端鉴定，我们对产甲烷八叠球菌嗜热蛋白酶 LysargiNase 进行了表征，该酶可在 55°C 以下切割赖氨酸和精氨酸。与胰蛋白酶不同，LysargiNase 生成的肽具有 N 端赖氨酸或精氨酸残基，并以 b 离子主导的谱图进行片段化。这提高了蛋白质 C 末端肽的鉴定和几个富含精氨酸的磷酸化位点的分配。值得注意的是，切割也发生在甲基化或二甲基化的赖氨酸和精氨酸上，有助于检测这些表观遗传修饰。
