On-tissue protein identification and imaging by MALDI-ion mobility mass spectrometry.
MALDI imaging mass spectrometry (MALDI-IMS) has become a powerful tool for the detection and localization of drugs, proteins, and lipids on-tissue. Nevertheless, this approach can only perform identification of low mass molecules as lipids, pharmaceuticals, and peptides. In this article, a combination of approaches for the detection and imaging of proteins and their identification directly on-tissue is described after tryptic digestion. Enzymatic digestion protocols for different kinds of tissues--formalin fixed paraffin embedded (FFPE) and frozen tissues--are combined with MALDI-ion mobility mass spectrometry (IM-MS). This combination enables localization and identification of proteins via their related digested peptides. In a number of cases, ion mobility separates isobaric ions that cannot be identified by conventional MALDI time-of-flight (TOF) mass spectrometry. The amount of detected peaks per measurement increases (versus conventional MALDI-TOF), which enables mass and time selected ion images and the identification of separated ions. These experiments demonstrate the feasibility of direct proteins identification by ion-mobility-TOF IMS from tissue. The tissue digestion combined with MALDI-IM-TOF-IMS approach allows a proteomics "bottom-up" strategy with different kinds of tissue samples, especially FFPE tissues conserved for a long time in hospital sample banks. The combination of IM with IMS marks the development of IMS approaches as real proteomic tools, which brings new perspectives to biological studies.
Stauber
Jonathan
J
FOM Institute for Atomic and Molecular Physics, Amsterdam, The Netherlands.
MacAleese
Luke
L
Franck
Julien
J
Claude
Emmanuelle
E
Snel
Marten
M
Kaletas
Basak Kükrer
BK
Wiel
Ingrid M V D
IM
Wisztorski
Maxence
M
Fournier
Isabelle
I
Heeren
Ron M A
RM
D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D008297:Male; D010449:Peptide Mapping / Q000379:methods*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D013048:Specimen Handling / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D046888:Tissue Array Analysis / Q000379:methods*; D014357:Trypsin / Q000737:chemistry
D020543:Proteome; D014357:Trypsin