title: Top-down-assisted bottom-up method for homologous protein sequencing: hemoglobin from 33 bird species.
abstract: Ticks are vectors for disease transmission because they are indiscriminant in their feeding on multiple vertebrate hosts, transmitting pathogens between their hosts. Identifying the hosts on which ticks have fed is important for disease prevention and intervention. We have previously shown that hemoglobin (Hb) remnants from a host on which a tick fed can be used to reveal the host's identity. For the present research, blood was collected from 33 bird species that are common in the U.S. as hosts for ticks but that have unknown Hb sequences. A top-down-assisted bottom-up mass spectrometry approach with a customized searching database, based on variability in known bird hemoglobin sequences, has been devised to facilitate fast and complete sequencing of hemoglobin from birds with unknown sequences. These hemoglobin sequences will be added to a hemoglobin database and used for tick host identification. The general approach has the potential to sequence any set of homologous proteins completely in a rapid manner. Graphical Abstract ᅟ.
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, 43210, USA.|Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, 85721, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, 85721, USA.
authors.forename: Ünige A
authors.initials: ÜA
authors.lastname: Laskay
authors.affiliation: Emerging and Acute Infectious Diseases Branch, Department of State Health Services, Austin, TX, 78756, USA.
authors.forename: Inger-Marie E
authors.initials: IM
authors.lastname: Vilcins
authors.affiliation: Microbiology and Molecular Genetics, Medicine, and Ecology and Evolutionary Biology, University of California, Irvine, CA, 92687, USA.
authors.forename: Alan G
authors.initials: AG
authors.lastname: Barbour
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, 43210, USA. wysocki.11@osu.edu.|Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, 85721, USA. wysocki.11@osu.edu.
authors.forename: Vicki H
authors.initials: VH
authors.lastname: Wysocki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000032:analysis / Q000737:chemistry / Q000145:classification; D001717:Birds; D030562:Databases, Protein; D006454:Hemoglobins / Q000032:analysis / Q000737:chemistry / Q000145:classification; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D020539:Sequence Analysis, Protein / Q000379:methods*; D012984:Software; D053719:Tandem Mass Spectrometry
chemical_list: D030161:Avian Proteins; D006454:Hemoglobins; D010446:Peptide Fragments
keywords: Bottom-up; Customized database; Hemoglobin; Sequencing; Top-down