Ouyang Zheng, Takáts Zoltán, Blake Thomas A, Gologan Bogdan, Guymon Andy J, Wiseman Justin M, Oliver Justin C, Davisson V Jo, Cooks R Graham
Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA.
Science. 2003 Sep 5;301(5638):1351-4. doi: 10.1126/science.1088776. Epub 2003 Aug 14.
Intact, multiply protonated proteins of particular mass and charge were selected from ionized protein mixtures and gently landed at different positions on a surface to form a microarray. An array of cytochrome c, lysozyme, insulin, and apomyoglobin was generated, and the deposited proteins showed electrospray ionization mass spectra that matched those of the authentic compounds. Deposited lysozyme and trypsin retained their biological activity. Multiply charged ions of protein kinase A catalytic subunit and hexokinase were also soft-landed into glycerol-based liquid surfaces. These soft-landed kinases phosphorylated LRRASLG oligopeptide and D-fructose, respectively.
从离子化的蛋白质混合物中选择特定质量和电荷的完整多质子化蛋白质，并将其轻柔地落在表面的不同位置以形成微阵列。生成了细胞色素c、溶菌酶、胰岛素和脱辅基肌红蛋白的阵列，沉积的蛋白质显示出与真实化合物相匹配的电喷雾电离质谱。沉积的溶菌酶和胰蛋白酶保留了它们的生物活性。蛋白激酶A催化亚基和己糖激酶的多电荷离子也被软着陆到基于甘油的液体表面。这些软着陆的激酶分别使LRRASLG寡肽和D-果糖磷酸化。
