Mass spectrometric detection of affinity purified crosslinked peptides
Mass spectrometric detection of affinity purified crosslinked peptides
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DOI：
10.1016/s1044-0305(04)00154-0
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发表时间：
2004-06-01
影响因子：
3.2
通讯作者：
Kennel, SJ
中科院分区：
文献类型：
--
作者：
Hurst, GB;Lankford, TK;Kennel, SJ
Chemical crosslinking of proteins combined with mass spectrometric analysis of the tryptic digest of the products shows considerable promise as a tool for interrogating structure and geometry of proteins and protein complexes. An impediment to the use of this tool has been the difficulty of distinguishing crosslinked peptide pairs from non-crosslinked peptides, and from the products of side reactions. We describe the use of a commercially available biotinylated crosslinking reagent, sulfo-SBED, that allows affinity-based enrichment of crosslinked species. An intramolecular crosslink is prepared using the peptide neurotensin as a model system. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra show the predicted crosslinking product, as well as several side products. Finally, we describe the optimized enrichment of biotinylated species, and reduction of non-specific binding, for a batch-mode affinity separation based on immobilized monomeric avidin. (C) 2004 American Society for Mass Spectrometry.
