Automated 16plex plasma proteomics with real-time search and ion mobility mass spectrometry enables large scale profiling in naked mole-rats and mice.

ABSTRACT: Performing large-scale plasma proteome profiling is challenging due to limitations imposed by lengthy preparation and instrument time. We present a fully Automated Multiplexed Proteome Profiling Platform (AutoMP3) using the Hamilton VantageTM liquid handling robot capable of preparing hundreds to thousands of samples. To maximize protein depth in single shot runs we combined 16plex Tandem Mass Tags (TMTpro) with high-field asymmetric waveform ion mobility spectrometry (FAIMS Pro) and real-time search (RTS). We quantified over 40 proteins / min / sample, doubling the previously published rates. We applied AutoMP3 to investigate the naked mole-rat plasma proteome both as a function of circadian cycle and in response to ultraviolet (UV) treatment. In keeping with the lack of synchronized circadian rhythms in naked mole-rats, we find few circadian patterns in plasma proteins over the course of 48hr. Furthermore, we quantify many disparate changes between mice and naked mole-rats at both 48hr and one week after UV exposure. These species differences in plasma protein temporal responses could contribute to the pronounced cancer resistance observed in naked mole-rats.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Heterocephalus Glaber Mus Musculus (mouse)

TISSUE(S): Blood Plasma

SUBMITTER: Sasha Gaun  

LAB HEAD: Fiona E. McAllister

PROVIDER: PXD022891 | Pride | 2021-02-04

REPOSITORIES: Pride