Publications - Cell and Metabolic Biology
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This page was last modified on 27 Jan 2025 .
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Publications - Cell and Metabolic Biology
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Mono‐ and poly‐adenosine diphosphate (ADP)‐ribosylation are common post‐translational modifications incorporated by sequence‐specific enzymes at, predominantly, arginine, asparagine, glutamic acid or aspartic acid residues, whereas non‐enzymatic ADP‐ribosylation (glycation) modifies lysine and cysteine residues. These glycated proteins and peptides (Amadori‐compounds) are commonly found in organisms, but have so far not been investigated to any great degree. In this study, we have analyzed their fragmentation characteristics using different mass spectrometry (MS) techniques. In matrix‐assisted laser desorption/ionization (MALDI)‐MS, the ADP‐ribosyl group was cleaved, almost completely, at the pyrophosphate bond by in‐source decay. In contrast, this cleavage was very weak in electrospray ionization (ESI)‐MS. The same fragmentation site also dominated the MALDI‐PSD (post‐source decay) and ESI‐CID (collision‐induced dissociation) mass spectra. The remaining phospho‐ribosyl group (formed by the loss of adenosine monophosphate) was stable, providing a direct and reliable identification of the modification site via the b‐ and y‐ion series. Cleavage of the ADP‐ribose pyrophosphate bond under CID conditions gives access to both neutral loss (347.10 u) and precursor‐ion scans (m/z 348.08), and thereby permits the identification of ADP‐ribosylated peptides in complex mixtures with high sensitivity and specificity. With electron transfer dissociation (ETD), the ADP‐ribosyl group was stable, providing ADP‐ribosylated c‐ and z‐ions, and thus allowing reliable sequence analyses.
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The pigments of Opuntia ficus‐indica fruits, which are derived from the betalain rather than anthocyanin pathway, have an extraordinary range in colour from lime green, orange, red to purple. This is a result from varying concentrations and proportions of about half a dozen betaxanthins and betacyanins. The yellow‐orange betaxanthins are derived from spontaneous condensation of betalamic acid with amines or amino acids. The reddish‐purple betacyanins are enzymatically formed from betalamic acid and cyclo ‐dihydroxyphenylalanine (DOPA) yielding betanidin and further glycosylated on either of the two hydroxyls of the cyclo ‐DOPA moiety. In the present work, degenerated primers were used to obtain partial genomic sequences of two major genes in the biosynthetic pathway for betalains, that is the 4,5‐extradiol dioxygenase which forms the betalamic acid responsible for the yellow colour and a putative 5‐O ‐glucosyltransferase which glycosylates betanidin in Dorotheanthus bellidiformis and may be responsible for the red colour. Differences in the genomic DNA between coloured versus non‐coloured varieties were not found. Regulatory mechanisms seem to independently control pigmentation of O. ficus‐indica fruit tissues for inner core, peel and epidermis. Core pigmentation occurs first and well before fruit maturity and peel pigmentation. Peel pigmentation is fully developed at maturity, presumably related to maximum soluble solids. Epidermal pigmentation appears to be independent of core and peel pigmentation, perhaps because of light stimulation. Similar control mechanisms exist through transcription factors for the major enzyme regulating anthocyanin production in grapes.
This page was last modified on 27 Jan 2025 .