Mass Spectrometry and Antibody-Based Characterization of Blood Vessels from Brachylophosaurus canadensis
Timothy P. Cleland*†, Elena R. Schroeter‡, Leonid Zamdborg§, Wenxia Zheng‡, Ji Eun Lee§∥, John C. Tran⊥, Marshall Bern#, Michael B. Duncan∇○, Valerie S. Lebleu∇○◆, Dorothy R. Ahlf⊥, Paul M. Thomas⊥, Raghu Kalluri∇○◆¶, Neil L. Kelleher⊥, and Mary H. Schweitzer‡□
†Marine, Earth and Atmospheric Sciences, ‡Department of Biological Sciences, North Carolina State University, Raleigh, North Carolina 27695, United States
§ Department of Chemistry, University of Illinois, Urbana, Illinois 61801, United States
∥ Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea
⊥ Departments of Chemistry, Molecular Biosciences and the Proteomics Center of Excellence, Northwestern University, Evanston, Illinois 60208, United States
# Protein Metrics, San Carlos, California 94070, United States
∇ Division of Matrix Biology, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02115, United States
○ Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115, United States
◆ Department of Cancer Biology, Metastasis Research Center, University of Texas MD Anderson Cancer Center, Houston, Texas 77054, United States
¶ Department of Biological Chemistry and Molecular Pharmacology and Harvard-MIT Division of Health Sciences and Technology, Harvard University, Cambridge, Massachusetts 02139, United States
□ North Carolina Museum of Natural Sciences, Raleigh, North Carolina 27601, United States
J. Proteome Res., Article ASAP
DOI: 10.1021/acs.jproteome.5b00675
Publication Date (Web): November 23, 2015
Copyright © 2015 American Chemical Society
*E-mail: tpcleland@utexas.edu. Phone: +1-919-515-7838.
Abstract
Structures similar to blood vessels in location, morphology, flexibility, and transparency have been recovered after demineralization of multiple dinosaur cortical bone fragments from multiple specimens, some of which are as old as 80 Ma. These structures were hypothesized to be either endogenous to the bone (i.e., of vascular origin) or the result of biofilm colonizing the empty osteonal network after degradation of original organic components. Here, we test the hypothesis that these structures are endogenous and thus retain proteins in common with extant archosaur blood vessels that can be detected with high-resolution mass spectrometry and confirmed by immunofluorescence. Two lines of evidence support this hypothesis. First, peptide sequencing of Brachylophosaurus canadensis blood vessel extracts is consistent with peptides comprising extant archosaurian blood vessels and is not consistent with a bacterial, cellular slime mold, or fungal origin. Second, proteins identified by mass spectrometry can be localized to the tissues using antibodies specific to these proteins, validating their identity. Data are available via ProteomeXchange with identifier PXD001738.
Keywords: Brachylophosaurus canadensis; blood vessels; dinosaur; cytoskeleton; actin; tubulin; myosin; tropomyosin; taphonomy; preservation
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