The ability to obtain molecular sequence for extinct organisms will have a dramatic impact on the assessment of the evolution of species. Ancient fossils from extinct organisms are usually degraded and little organic material remains for molecular analysis. We are developing a very sensitive approach using mass spectrometry and bioinformatics for extracting and sequencing available protein (mainly collagen) that remains in well-preserved bones. For fossils as old as 68 million years such as a Tyrannosaurus rex preserving soft tissues, we have extracted and sequenced small fragments of collagen as well as acquired significant sequence information for a 160,000-600,000 year old mastodon fossil. For such specimens, DNA is not readily available and protein is the only sequenceable organic material. These sequences were acquired in low resolution so that many isobaric amino acids and modified amino acids could not be sufficiently resolved. Funding from this Major Research Instrumentation (MRI) Program grant provides partial support for the acquisition of a ThermoScientific LTQ-Orbitrap mass spectrometer capable of ultra high resolution and mass accuracy that will afford the PIs the ability to sufficiently resolve these sequences and their modifications without ambiguity and with greater sensitivity to acquire sequence information on fossils that are many millions of years old. Many fossils showing excellent preservation fossils have been recently excavated and these will be sequenced using the PIs approach. These sequences will then be used to assess their evolutionary relationships with modern organisms as well as other extinct organisms. The instrument will be housed at Beth Israel Deaconess Medical Center and 33% of the instrument time will be dedicated for terrestrial research. Students from North Carolina State University conducting research in vertebrate and molecular paleontology will be afforded the opportunity for training on this state-of-the-art technology.
