title: Cardiac extracellular proteome profiling and membrane topology analysis using glycoproteomics.
abstract: OBJECTIVE
  Extracellular proteins are easily accessible, which presents a subproteome of molecular targets that have high diagnostic and therapeutic potential. Efforts have been made to catalog the cardiac extracellular matridome and analyze the topology of identified proteins for the design of therapeutic targets. Although many bioinformatics tools have been developed to predict protein topology, topology has been experimentally validated for only a very small portion of membrane proteins. The aim of this study was to use a glycoproteomics and MS approach to identify glycoproteins in the extracellular matridome of the infarcted left ventricle (LV) and provide experimental evidence for topological determination.
  
  
  METHODS
  Glycoproteomics analysis was performed on eight biological replicates of LV samples from wild-type mice at 7 days following myocardial infarction using SPE of glycopeptides, followed by mass spectrometric identification of N-linked glycosylation sites for topology assessment.
  
  
  RESULTS
  We identified hundreds of glycoproteins, and the identified N-glycosylation sites provide novel information on the correct topology for membrane proteins present in the infarct setting.
  
  
  CONCLUSIONS
  Our data provide the foundation for future studies of the LV infarct extracellular matridome, which may facilitate the discovery of drug targets and biomarkers.
authors.affiliation: San Antonio Cardiovascular Proteomics Center, San Antonio, TX, USA; Mississippi Center for Heart Research, Department of Physiology and Biophysics, University of Mississippi Medical Center, Jackson, MS, USA; Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tian
authors.forename: Tejaswi
authors.initials: T
authors.lastname: Koganti
authors.forename: Zhihao
authors.initials: Z
authors.lastname: Yao
authors.forename: Presley
authors.initials: P
authors.lastname: Cannon
authors.forename: Punit
authors.initials: P
authors.lastname: Shah
authors.forename: Laura
authors.initials: L
authors.lastname: Pietrovito
authors.forename: Alessandra
authors.initials: A
authors.lastname: Modesti
authors.forename: Paul
authors.initials: P
authors.lastname: Aiyetan
authors.forename: Kristine
authors.initials: K
authors.lastname: DeLeon-Pennell
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Ma
authors.forename: Ganesh V
authors.initials: GV
authors.lastname: Halade
authors.forename: Chindo
authors.initials: C
authors.lastname: Hicks
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Merry L
authors.initials: ML
authors.lastname: Lindsey
mesh_terms: D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism*; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006352:Heart Ventricles / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009203:Myocardial Infarction / Q000378:metabolism / Q000473:pathology; D040901:Proteomics
chemical_list: D006023:Glycoproteins
keywords: Extracellular matridome; Glycoprotein; Left ventricle; Matrix metalloproteinase; Membrane orientation; Myocardial infarction