title: Proteomic profiling of a snake venom using high mass detection MALDI-TOF mass spectrometry.
abstract: Proteomic profiling involves identification and quantification of protein components in complex biological systems. Most of the mass profiling studies performed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) have been restricted to peptides and small proteins (<20 kDa) because the sensitivity of the standard ion detectors decreases with increasing ion mass. Here we perform a protein profiling study of the snake venom Sistrurus miliarius barbouri, comparing 2D gel electrophoresis and reversed-phase high-performance liquid chromatography (HPLC) with a high mass cryodetector MALDI-TOF instrument (Macromizer), whose detector displays an uniform sensitivity with mass. Our results show that such MS approach can render superior analysis of protein complexity compared with that obtained with the electrophoretic and chromatographic approaches. The summation of ion impacts allows relative quantification of different proteins, and the number of ion counts correlates with the peak areas in the reversed-phase HPLC. Furthermore, the sensitivity reached with the high mass cryodetection MS technology clearly exceeds the detection limit of standard high-sensitivity staining methods.
authors.affiliation: Institut de Biotecnologia i Biomedicina and Departament de Bioquimica i Biol. Mol., Universitat Autònoma de Barcelona, Bellaterra (Barcelona), Spain.
authors.forename: Oscar
authors.initials: O
authors.lastname: Yanes
authors.forename: Francesc X
authors.initials: FX
authors.lastname: Avilés
authors.forename: Ryan
authors.initials: R
authors.lastname: Wenzel
authors.forename: Alexis
authors.initials: A
authors.lastname: Nazabal
authors.forename: Renato
authors.initials: R
authors.lastname: Zenobi
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D017819:Viperidae
chemical_list: D003435:Crotalid Venoms