Fig. 6

Biochemical characterization of 34 AID orthologs from species related closely to the Atlantic cod. A The left panel shows a species tree adapted from Malmstrom et al. [33]. AID proteins from species colored blue were synthesized in the lab to study their biochemical properties. Channel catfish and human AIDs were also purified and tested. We could not find any aicda gene in the genomic sequence of B. cantori (colored in red). The right panel shows the primary sequence of the AID enzymes that were expressed and purified. B The optimal temperature profiles of extant AID orthologs was assessed using our standard alkaline cleavage assay and bub7TGC substrate. Two to three independent protein preparations of each AID ortholog were tested in duplicates. Results are plotted as deamination activity percentage. The incubation duration, minimum, and maximum temperature limits were tailored to the activity level of each purified AID obtained in the preliminary results. For better representation, results were graphed based on the AIDs’ activity level. A through D show AIDs with low to high activity levels. Data is graphed as mean ± SEM (n = 4). Abbreviations: Gm: Gadus morhua; Bs: Boreogadus saida; Ag: Arctogadus glacialis; Memrla: Merlangius merlangus; Ma: Melanogrammus aeglefinus; Pv: Pollachius virens; Ga: Gadiculus argenteus; Tmi: Trisopterus minutus; Bb: Brosme brosme; Mmol: Molva molva; Llo: Lota lota; Pp: Phycis phycis; Pb: Phycis blennoides; Mo: Malacocephalus occidentalis; Mb: Macrourus berglax; Bm: Bathygadus melanobranchus; Lla: Laemonema laureysi; Mmor: Mora mora; Tmu: Trachyrincus murrayi; Tsc: T. scabrous; Mma: Muraenolepis marmoratus; Mz: Melanonus zugmayeri; Mmerlu: Merluccius merluccius; Sc: Stylepnorus chordates; Cr: Cyttopsis roseus; Zf: Zeus faber; Tsu: Typhlichthys subterraneus; Pt: Percopsis transmontane; Pj: Polymixia japonica; Ss: Salmo salar; Dr: Danio rerio; Ol: Oryzias latipes; Tr: Takifugu rubripes; Ip: Ictalurus punctatus; Hs: Homo sapiens