title: Homogenization of tissues via picosecond-infrared laser (PIRL) ablation: Giving a closer view on the in-vivo composition of protein species as compared to mechanical homogenization.
abstract: Posttranslational modifications and proteolytic processing regulate almost all physiological processes. Dysregulation can potentially result in pathologic protein species causing diseases. Thus, tissue species proteomes of diseased individuals provide diagnostic information. Since the composition of tissue proteomes can rapidly change during tissue homogenization by the action of enzymes released from their compartments, disease specific protein species patterns can vanish. Recently, we described a novel, ultrafast and soft method for cold vaporization of tissue via desorption by impulsive vibrational excitation (DIVE) using a picosecond-infrared-laser (PIRL). Given that DIVE extraction may provide improved access to the original composition of protein species in tissues, we compared the proteome composition of tissue protein homogenates after DIVE homogenization with conventional homogenizations. A higher number of intact protein species was observed in DIVE homogenates. Due to the ultrafast transfer of proteins from tissues via gas phase into frozen condensates of the aerosols, intact protein species were exposed to a lesser extent to enzymatic degradation reactions compared with conventional protein extraction. In addition, total yield of the number of proteins is higher in DIVE homogenates, because they are very homogenous and contain almost no insoluble particles, allowing direct analysis with subsequent analytical methods without the necessity of centrifugation.
  
  
  CONCLUSIONS
  Enzymatic protein modifications during tissue homogenization are responsible for changes of the in-vivo protein species composition. Cold vaporization of tissues by PIRL-DIVE is comparable with taking a snapshot at the time of the laser irradiation of the dynamic changes that occur continuously under in-vivo conditions. At that time point all biomolecules are transferred into an aerosol, which is immediately frozen.
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Kwiatkowski
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Wurlitzer
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Krutilin
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Kiani
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Nimer
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Omidi
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Mannaa
authors.affiliation: Beiersdorf AG, Research & Development, Unnastrasse 48, 20245, Hamburg, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Bussmann
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Department of Tumor Biology, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Bartkowiak
authors.affiliation: Max Planck Institute for the Structure and Dynamics of Matter, Atomically Resolved Dynamics Division, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kruber
authors.affiliation: Max Planck Institute for the Structure and Dynamics of Matter, Atomically Resolved Dynamics Division, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Uschold
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Steffen
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Lübberstedt
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Küpker
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Department of Otorhinolaryngology, Head and Neck Surgery and Oncology, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Petersen
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Department of Otorhinolaryngology, Head and Neck Surgery and Oncology, Martinistraße 52, 20246 Hamburg, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Knecht
authors.affiliation: Max Planck Institute for the Structure and Dynamics of Matter, Atomically Resolved Dynamics Division, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: N O
authors.initials: NO
authors.lastname: Hansen
authors.affiliation: Techna Institute for the Advancement of Technology for Health, University Health Network, Toronto, ON M5G-1P5, Canada & Department of Medical Biophysics, University of Toronto, 101 College Street Suite 15-701, Toronto, ON M5G 1L7, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Zarrine-Afsar
authors.affiliation: Max Planck Institute for the Structure and Dynamics of Matter, Atomically Resolved Dynamics Division, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: W D
authors.initials: WD
authors.lastname: Robertson
authors.affiliation: Max Planck Institute for the Structure and Dynamics of Matter, Atomically Resolved Dynamics Division, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: R J D
authors.initials: RJD
authors.lastname: Miller
authors.affiliation: University Medical Centre Hamburg-Eppendorf, Institute for Clinical Chemistry, Department for Mass Spectrometric Proteomics, Martinistraße 52, 20246 Hamburg, Germany. Electronic address: hschluet@uke.de.
authors.forename: H
authors.initials: H
authors.lastname: Schlüter
mesh_terms: D000818:Animals; D006801:Humans; D007259:Infrared Rays; D007834:Lasers; D051379:Mice; D014066:Palatine Tonsil / Q000737:chemistry*; D010179:Pancreas / Q000737:chemistry*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods; D017208:Rats, Wistar; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods
keywords: Mass spectrometry; PIRL-DIVE; Protein species; Proteolysis; Tissue homogenization