Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-17177 Stockholm, Sweden.
Mol Cell Proteomics. 2011 Sep;10(9):M110.006510. doi: 10.1074/mcp.M110.006510. Epub 2011 May 24.
A membrane cell for hydrogen and deuterium exchange on-line with mass spectrometry has been developed to monitor protein-protein interactions and protein conformations. It consists of two channels separated by a semipermeable membrane, where one channel carries the protein sample and the other deuterium oxide. The membrane allows transfer of deuterium oxide into the sample flow. The labeling time is controlled via the flow rate in the sample channel. This cell was validated against three models commonly used in hydrogen-deuterium exchange mass spectrometry: monitoring of folded and unfolded states in a protein, mapping the protein secondary structure at the peptide level, and detection of protein and antibody interactions. The system avoids the conventionally used sample dilution and handling, allowing for potential automation.
一种用于与质谱联用的在线氢氘交换的膜片细胞已经被开发出来，以监测蛋白质-蛋白质相互作用和蛋白质构象。它由两个通道组成，中间由半透膜隔开，一个通道携带蛋白质样品，另一个通道携带重水。膜允许重水向样品流转移。标记时间通过样品通道中的流速来控制。该细胞与氢氘交换质谱中常用的三种模型进行了验证：监测蛋白质的折叠和未折叠状态、在肽水平上绘制蛋白质二级结构以及检测蛋白质和抗体相互作用。该系统避免了传统上使用的样品稀释和处理，允许潜在的自动化。
