Grams, A. Fleischer, K. Pawlak, M. Matczuk, A. Siwek, L. Xifeng, S. Aleksenko, A. Timerbaev, M. Jarosz Binding abilities of copper to phospholipides. Jakl, D. Peukert, A. Matros, H. Mock Expanding role of in vitro simulation of cytochrome P metabolism for drugs of abuse. Jakub Mielczarek, H. Raoof, J. Silberring Software for Pathway Informed Metabolomics. Thiemann Development of a spectral interconversion algorithm for desorption ionization techniques.
Golf, M. Walther-Schmidt, Z. Zehethofer, B. Kulle, C. Wewetzer, P. Holterhus, F. Lottspeich, Prof. Meyer, Dr.
Frochaux, M. Trusch, D. Hildebrand, M. Linscheid, H. Jankowska, M. Sosnowska, P. Karpowicz, K. Radziszewska, P. Stefanowicz, A. Schlosser, E. Haaf Specific detection of arginine-derived advanced glycation end-products and their localization in proteins.
Schmidt, D. Singer, R. Hoffmann, A. Pischetsrieder How to handle huge amount of mass spectrometric data? A new strategy with the Achroma software. Scheerle, C. Kaufmann, M. Krappmann, J. Grassmann, T.
Kot- -Wasik, A. Jakimska, A. Wasik, J. Zwiener, M. Daugherty, S. Koc- -Latoszek, J. Garcia-Reyes, A. Molina Diaz, M. Fiebig, H-G. Schmalz, M. Schadt, F. Porbeck, R. Almeida, S. Kallbach, J. Kirschbaum, T. Kraj, H. Brouwer, N. Reinhoud, P. Mielczarek, J. Silberring, J-P. Chervet High-resolution time-of-flight mass spectrometry using folded flight path technology for profiling of steroids and steroid metabolites in urine. Wendt, K.
Siek, J. Patrick, E. Guice Maldi-ToF Analysis in the study of luminescent polyethers and poluthioeters with the crabazolyl unit in the substituent for oled applications. Swinarew, J. Gabor, S. Golba, B. Pirkl, J. Soltwisch, F. Draude, K. Szewczuk, M. Rudowska, R. Cydzik, D. Wojewska, A.
Kluczyk, P. Session Chair: M. Driving this approach is the wide availability of mass spectrometers for analytical proteomics; these should also be applicable to protein footprinting. FPOP is fast owing to the use of free radicals; radical generation occurs in low ns, and the radical reactions are complete in 1 microsec. Only a single conformation of the protein exists during the footprinting. Besides OH radicals, other radicals can be used including the sulfate and carbonate radical anions.
FPOP can also be the probe in a classic, two-laser pump-probe experiment whereby a temperature-jump perturbation is produced as a pump by one laser and a second laser initiates FPOP as the probe.
Subsequent analysis is by MS. This experiment is capable of probing protein structural dynamics at the sub millisecond level with improved sensitivity and more detailed structural resolution than any physical chemical method.
Better understanding of folding will emerge from this approach, which is more informative thanthe usual physical probe methods e. At each time, aliquot was removed, quenched by lowering ph to 2. Samples infused through fused silica; the laser irradiated a window at a pulse frequency giving single-shot exposure. Other labeling strategies will be illustrated to understand the dimerization of Her4 protein in a membrane and its subsequent phosphorylation.
The results of a combination of these approaches will then be applied to understand the oligomerization of apoe. In recent years it has been demonstrated, however, that electrospray ionization can indeed provide a close link between observations in the idealized conditions of a mass spectrometer and real chemistry occurring in solution. Method: The experiments described use electrospray ionization ESI with a particular emphasis on concentration series and the effects of the concentration of the feed solution on the mass spectrometric data obtained.
Preliminary Data: The gas-phase chemistry of metal ions is briefly reviewed and a few examples for correlations between ESI-MS measurements and condensed phase properties. These include several solvated metal ions [1, 2] and metal-mediated coupling reactions [3, 4]. In addition to qualitative aspects concerning the type and intensities of certain species in solution, valuable mechanistic information can be achieved which bear relevance for preparative chemistry.
In fortunate cases, it is even possible to link the chemistry in solution directly with that occurring in the gas phase and also in the transfer between the condensed and gaseous state .
In this context, we also propose the first quantitative correlations between gas-phase measurements and condensed-phase properties [2, 5]. Novel Aspects: Quantitative correlations between gas-phase and condensed-phase properties.
Tsierkezos, J. Karwowska, K. Fijalkowski, M. Agrawal, D. Svec, M. The basis of the HDX-MS method is that backbone amide hydrogens that are engaged in stable hydrogen bonds in the folded conformation are protected against exchange with the solvent. Structural fluctuations and local unfolding events that transiently break these hydrogen bonds cause exposure of the amide hydrogens to the solvent and this leads to exchange.
The exchange kinetics of amide backbone hydrogens is therefore a direct probe for the structural dynamics of the protein backbone. Method: When a protein is incubated in D 2 O, its global deuterium uptake is readily monitored by mass spectrometry as each incorporated deuteron causes a mass increase of 1 Da. To obtain local information on the deuterium incorporation, the labeled protein is typically digested by pepsin at cold acidic conditions where the amide hydrogen exchange reaction is quenched i.
Preliminary Data: With this bottom-up approach, we have investigated the structural dynamics of proteases and protease-inhibitors implicated in extracellular proteolysis, kinases and allosteric activation mechanisms. The spatial resolution of the bottom-up approach was until recently on the peptide level. We have recently demonstrated the utility of gas-phase electron-based fragmentation to obtain site-specific deuterium levels in peptides. Following its development in and its commercial introduction in , the DART ion source has been applied to an extraordinarily broad range of applications.
This may be attributed to the accessibility of the atmospheric-pressure ionization region and the relative simplicity of the mass spectra that are the end result of a set of complex reaction mechanisms.
Method: Recently, we have operated the DART with a relatively high gas temperature in a pyrolysis mode for the identification of wood samples. The first such application was to distinguish red oak Quercus rubra and white oak Quercus alba.
Chemometric tools were applied to the negative-ion DART mass spectra for classification. This approach was subsequently used by the US Fish and Wildlife Service to identify Brazilian Rosewood, which is an endangered species that is banned for importation. The unexpectedly high accuracy may result from the use of a relatively large set of biomarker ions. Preliminary Data: Other recent applications of DART have included the identification of so-called spice designer drugs, detection of condom lubricants in fingerprints, and screening for banned pesticides in imported fruit.
The latter application makes use of DART in transmission mode, providing more reproducible sampling and improved quantitation. A novel variation of transmission-mode DART involves spotting samples on wire mesh on a disposable card. The mesh has a resistive area that can be heated rapidly in the DART gas stream by passing an electrical current through the mesh.
This simplifies and speeds analysis by eliminating such variables as sample placement and DART gas temperature. This has previously been demonstrated for the identification of melamine contamination in pet food and powdered milk. The information content of the generated MS images greatly depends on the quality of the underlying mass spectral data. We have recently introduced a method that combines the high performance features of Fourier transform mass spectrometers with a spatial resolution in the low micrometer range .
We have now optimized our workflow for the Exactive Orbitrap and applied it to the detailed analysis of phospholipids in a number of biological samples. In addition data processing and analysis strategies have been expanded and optimized.
Method: Sections of mammalian, insect and plant tissue were prepared with a cryomicrotome. Matrix was applied by a home-built spraying device . MS imaging experiments were performed with a home-built atmospheric-pressure imaging source  attached to an Exactive Orbitrap mass spectrometer Thermo Scientific GmbH, Bremen.
Mass accuracy was better than 2 ppm root mean square under imaging conditions, i. In all experiments the high resolution and mass accuracy proved to be essential for specific image generation and reliable identification of analytes. Preliminary Data: A dedicated sample preparation protocol was established for the analysis of single cells.
We were able for the first time to identify larger metabolites phospholipids and investigate their spatial distribution within native single cells in one measurement. Intact phospholipids and their acyl chain fragments were detected simultaneously in all ion fragmentation experiments in a human brain tumor sample. This allowed the tentative differentiation of isomeric lipid structures throughout the whole section within one experiment.
A complete set of positive and negative ion images was obtained simultaneously by periodically switching the polarity of the ion optics throughout the imaging experiment. This significantly increased the number of lipids that could be identified in a single experiment in mammalian tissue and thus improves the differentiation of tissue types. The effect of microbial infection on plants was studied based on changes in the phospholipid profiles.
Detailed distributions of phospholipids were detected within a root section of less than 1 mm in diameter. Whole body-sections of insects e. Phospholipid patterns were used to obtain morphological information and to assign signals of interest e. Statistical analysis tools including PCA and LDA were adopted and applied for semiautomic assignment of tissue types in mouse and human tissue sections.
Additional data analysis tools were made accessible by conversion to the common data format for MS imaging imzml The presented workflow with its improvements in sample preparation, measurement parameters and data processing significantly improves the biologically relevant information that can be obtained by mass spectrometry imaging.
Novel Aspects: Highly reliable biological information from MS imaging for mammalian, insect and plant samples. RAGE is an important mediator of the proinflammatory response involved in diverse pathophysiological states such as neurological disorders, stroke,amyloidosis, immune response, diabetes and inflammatory disorders, infectious disease and tumors.
It is a potential therapeutic target, but in spite of significant effort its structure, ligand binding mode and signal transduction pathway has not been characterised yet on the molecular level. Method: In the presented work we used hydrogen-deuterium exchange and mass spectrometry to gain insight into the structural properties of exrage the full extracellular part of the protein containing V, C1 and C2 domains and the structural consequences of its oligomerisation. Preliminary Data: The highlight of the present study is the intertwining of protected and fully exposed regions of the protein underscoring a highly dynamic character of large portions of this protein.
However, we show that upon oligomerisation the C1-C2 domain linker region, fully flexible in the monomer, becomes stabilised. Mass spectrometry based on electrospray ionization can be a useful tool for establishing the non-covalent complex stoichiometry as well as origin of conformational stabilization.
However, desolvation is a crucial step for maintaining at least partially the system conformation preformed in solution. Thus, under such careful conditions, ESI mass spectra can reflect partially the properties of survivor charged complexes in gas phase. On the other hand, from naked multiply charged systems solvent-free , the ion activation can provide some insights on the interaction sites on the folded molecule or in complex ions.
For reaching such information, a veritable arsenal is helpful for ion activation. Indeed, ergodic e. The presented studies will different illustrations of application with its limitation and the risk of conclusive mistake. From the latter, a heated hollow cathode operating at a current of 1. Double resonance experiments were used for ejection of selected fragment ions which could be intermediate to formation of a second generation product ion.
Preliminary results: From large monomeric systems as peptides, it is shown how their conformation, as well as their rigidity may influence dramatically the decomposition pathways occurring from the excited multicharged species.
The ECD process for positive species as well as EDD for negative ones leads to some differentiation of conformers and topoisomercompared to the behaviour observed by CID.
On the other hand, from various covalent and non-covalent ssdna and dsdna systems associated with drugs and peptides were investigated by using different activation modes according to the charge state and size, as well as the ion polarity.
It emerges from many results, a similar trend which characterizesion orientation dissociations. This appears to be specific to the used activation. From several examples, it seems, that independently of the charge polarity, the formation of salt bridge plays an important role in the conformational stabilization.
This stabilisation can be evidenced by cleavage of covalent bond rather than the salt bridge. Indeed, the previous thermochemical terms decrease strongly with the increasing of charge state.
During all the gas phase operations: i aggregate formation and ii desolvation steps, it seems that conformation like to solution can survive at least partially, by the increasing of electrostatic interactions during the disappearance of hydrophobic interactions. Indeed, the neighboring groups which interact in solution can be maintained closer by the electrostatic effects if already they pre-exist in solution, even relatively distant due to solvent.
Non-hydrocarbon compounds present in a crude oil are among the most deleterious to refining catalysts and confer adverse stability properties. The complexity of the individual petroleum fractions is quite high, even after initial fractionations i.
SARA , therefore the coupling of chromatography with an ultra-high resolution mass spectrometer is a successful choice. On the other hand, nitrogen aromatic heterocyclic fractions were separated as well on a polar aminocyano-bonded silica HPLC column PAC . This separation within the same class gives more detailed information about nitrogen compounds in crude oil samples. The column was kept at room temperature, and was connected to a UV diode array detector set at nm. Offline samples were collected via Nanomate autosampler, whereas a micro-splitter was used to control the flow rate for online measurements.
Preliminary Data: Normal-phase chromatography was used as the separation method, because the majority of the compounds of interest in this case are of lower polarity.
Using a mobile phase system consisting of n-hexane and isopropanol leads to better resolved chromatograms. A fraction of both peaks were collected offline every 20 seconds and by using a nano-esi Advion Nanomate. The N-selective separation was optimized into two major peaks: one broad peak appears after about one minute, and another peak elutes at around 20 minutes.
The first peak comprises mainly of hydrocarbons, whereas the second peak contains exclusively nitrogen species. ESI as an ion source was not adequate for this kind of investigation, when using n-hexane as an eluent. On the contrary, APCI measurements deliver good results, because it is an ideal method of ionization for low- to medium- polar compounds.
A relative wide spectrum of polar species was observed using APPI as an ion source, However the lack of using a dopant was obvious on the outcome of the results. On the other hand APLI can deliver efficient ionization of poly aromatic species of the non-polar peak detected at earlier retention times.
Increase of aromaticity of hydrocarbons and sulfur species in course of time was observed in the APLI measurement, in addition to the characterization of two distinct sulfur peaks in the non-polar fraction.
Bioanal Chem. A ,  Li M. The investigations have shown that this double loss is a general feature of the rhodamine B structure and that the fragmentation reaction takes place in the in the sidechain subsitution of the nitrogen atoms. This work shows the extension of these first experiments to molecules containing methyl and ethyl groups in different combinations, including deuteration.
The properties of the fragment spectra are interpreted towards their meaning for the underlying reaction mechanism. Method: The substances under investigation all had the basic structure of decarboxy-rhodamine B. They were synthesized using literature procedures [2, 3]. The experiments were conducted on a 9. The analytes were transferred into the gas phase via the electrospray technique.
Fragmentation was induced inside the ICR cell by two methods. First, collisional activation was performed using the SORI method and argon gas as collision partner. Additionally, a cleavage of CH 3 could be detected. This shows that, in this case, radicals are involved in the main losses and that these radicals can be seen on the timescale of the experiments.
Calculations of the reaction regarding a concerted mechanism on the one hand and a stepwise radical one on the other hand gave further insight to these findings. This explains why this first intermediate is accessible during the reaction.
Further measurements were performed with a substance carrying only one ethyl group at each nitrogen atom. Among the occurring peaks, the most interesting ones are due to the loss of CH 3 and C 2 H 5. They are higher in intensity than the respective even electron losses. Furthermore, the spectra show a peak that results from a combined cleavage of CH 3 and C 2 H 5. This shows that radicals in this system are far more likely to occur than in the ones investigated before.
Here the proposed mechanism for a concerted reaction cannot be realized, which makes the radical losses that intense. The latter shows, that a non-concerted component is present here. Novel Aspects: Laser and collision induced fragmentation of deuterated and non-deuterated xanthene dyes containing methyl and ethyl substituents.
Peters and, J. Grotemeyer, Rapid Commun. Mass Spectrom. Kimachi, K. Tanaka, F. Defining the relevant molecular mechanisms mediating pheromone signaling is an important step to manipulate pheromone-induced behaviors in pathogenic or agriculturally important pests. The only volatile pheromone identified in Drosophila is cis-vaccenyl acetate VA , a male-specific lipid that mediates aggregation behavior.
VA activates a few dozen olfactory neurons located in T1 sensilla on the antenna of both male and female flies. Here, we identify a neuronal receptor required for VA sensitivity. We identified two mutants lacking functional T1 sensilla and show that the expression of the VA receptor is dramatically reduced or eliminated.Preliminary Data: Here we Write awesome cover letter a calibration protocol for O, its global deuterium uptake is readily monitored by carbohydrates increase of 1 Da. Method: When Cis protein is incubated in D 2 TW-IMS syntheses using a series of naturally occurring glycan-type mass spectrometry as each incorporated acetate causes a mass. This simplifies and speeds analysis by eliminating such variables as sample placement and DART gas temperature. FPOP can also be the probe in a classic, two-laser pump-probe experiment whereby a temperature-jump perturbation is produced as a pump by one laser and a second laser initiates FPOP as the probe.
Method: When a protein is incubated in D 2 O, its global deuterium uptake is readily monitored by mass spectrometry as each incorporated deuteron causes a mass increase of 1 Da. A fraction of both peaks were collected offline every 20 seconds and by using a nano-esi Advion Nanomate. Waters Gmbh, Austria 5. Gel lanes were sliced into 11 pieces which were processed for In-gel digestion .
Preliminary results: From large monomeric systems as peptides, it is shown how their conformation, as well as their rigidity may influence dramatically the decomposition pathways occurring from the excited multicharged species. Grams, A. Ingendoh, M. Among the occurring peaks, the most interesting ones are due to the loss of CH 3 and C 2 H 5. Fleischer, K. Nakade, S.
In this context, we also propose the first quantitative correlations between gas-phase measurements and condensed-phase properties [2, 5]. Protein quantification across hundreds of experimental conditions. Among the occurring peaks, the most interesting ones are due to the loss of CH 3 and C 2 H 5. Meyer, Dr. Hoffmann, A. This shows that radicals in this system are far more likely to occur than in the ones investigated before.
Adamus, M. Singer, R. They are higher in intensity than the respective even electron losses.