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Although bottom-up proteomics using tryptic digests is widely used to locate post-translational modifications (PTM) in proteins, there are cases where the protein has several potential modification sites within a tryptic fragment and MS2 strategies fail to pinpoint the location. We report here a method using two proteolytic enzymes, trypsin and pepsin, in combination followed by tandem mass spectrometric analysis to provide fragments that allow one to locate the modification sites. We used this strategy to find a glycosylation site on bovine trypsin expressed in maize (TrypZean). Several glycans are present, and all are attached to a nonconsensus N-glycosylation site on the protein. 10.1021/ac1020722

Summary: There is limited information about the relative composition and health benefits of various honey consumed across Africa. This study aimed at estimating the bioactive constituents, in vitro radical scavenging and antibacterial activities of 16 kinds of honey obtained from different geographical locations in Kenya. Manuka 5 + honey was included for comparison. Some of the tested honey had biochemicals and bioactivities similar to or higher than Manuka 5 + honey. The honey exhibited DPPH radical scavenging ability, with several types of honey showing superior scavenging potential than Manuka 5 + honey, owing to their high phenol content. All types of honey inhibited the growth of E. coli and further showed a substantial amount of nonperoxide antimicrobial activity. The geographical origin of honey had an influence on its bioactive contents. Overall, these findings suggest that Kenyan honey has great therapeutic potential, and thus, its clinical application should not be overlooked. [ABSTRACT FROM AUTHOR]