title: Mass Spectrometry and Antibody-Based Characterization of Blood Vessels from Brachylophosaurus canadensis.
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.
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Cleland
authors.forename: Elena R
authors.initials: ER
authors.lastname: Schroeter
authors.affiliation: Department of Chemistry, University of Illinois , Urbana, Illinois 61801, United States.
authors.forename: Leonid
authors.initials: L
authors.lastname: Zamdborg
authors.forename: Wenxia
authors.initials: W
authors.lastname: Zheng
authors.affiliation: 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.
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Lee
authors.affiliation: Departments of Chemistry, Molecular Biosciences and the Proteomics Center of Excellence, Northwestern University , Evanston, Illinois 60208, United States.
authors.forename: John C
authors.initials: JC
authors.lastname: Tran
authors.affiliation: Protein Metrics , San Carlos, California 94070, United States.
authors.forename: Marshall
authors.initials: M
authors.lastname: Bern
authors.affiliation: 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.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Duncan
authors.affiliation: 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.
authors.forename: Valerie S
authors.initials: VS
authors.lastname: Lebleu
authors.affiliation: Departments of Chemistry, Molecular Biosciences and the Proteomics Center of Excellence, Northwestern University , Evanston, Illinois 60208, United States.
authors.forename: Dorothy R
authors.initials: DR
authors.lastname: Ahlf
authors.affiliation: Departments of Chemistry, Molecular Biosciences and the Proteomics Center of Excellence, Northwestern University , Evanston, Illinois 60208, United States.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Thomas
authors.affiliation: 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.
authors.forename: Raghu
authors.initials: R
authors.lastname: Kalluri
authors.affiliation: Departments of Chemistry, Molecular Biosciences and the Proteomics Center of Excellence, Northwestern University , Evanston, Illinois 60208, United States.
authors.forename: Neil L
authors.initials: NL
authors.lastname: Kelleher
authors.affiliation: North Carolina Museum of Natural Sciences , Raleigh, North Carolina 27601, United States.
authors.forename: Mary H
authors.initials: MH
authors.lastname: Schweitzer
mesh_terms: D000199:Actins / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001808:Blood Vessels / Q000033:anatomy & histology* / Q000378:metabolism* / Q000382:microbiology; D001842:Bone and Bones / Q000098:blood supply; D002645:Chickens; D025061:Dinosaurs / Q000033:anatomy & histology* / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique / Q000379:methods; D005580:Fossils / Q000033:anatomy & histology*; D013058:Mass Spectrometry; D008954:Models, Biological; D008969:Molecular Sequence Data; D009218:Myosins / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D040901:Proteomics / Q000379:methods; D016415:Sequence Alignment; D013045:Species Specificity; D020310:Struthioniformes; D014335:Tropomyosin / Q000235:genetics / Q000302:isolation & purification; D014404:Tubulin / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000199:Actins; D014335:Tropomyosin; D014404:Tubulin; D009218:Myosins
keywords: Brachylophosaurus canadensis; actin; blood vessels; cytoskeleton; dinosaur; myosin; preservation; taphonomy; tropomyosin; tubulin