Volume 2, #2, 2010

NOVA

TECHNICAL NOTES

Protein methodologies access acid-labile histidine phosphorylationUlli-Martin Hohenester, Karin Ludwig, Josef Krieglstein, Susanne Klumpp, Simone Königpp 71-76Abstract:Phosphohistidine is estimated to be two to three orders of magnitude more abundant in nature than phosphotyrosine but its bioanalytical detection is much less developed. Histidine phosphorylation is sensitive to acid which is one reason why it eluded common analysis strategies in the past. We have chemically phosphorylated a number of His-containing peptides with the purpose of elucidating chromatographic and mass spectrometric parameters for the analysis of phosphorylation sites. We found that typical proteomic methods for peptide studies including low-pH reversed phase chromatography and collision-induced dissociation can be applied. The half-life of the modification is in the order of several hours in acidic or formic acid-containing solvents. In tandem mass spectrometry it is dependent on peptide size, charge, sequence and experimental parameters if the phosphate is retained on backbone fragments or lost from the parent ion.A surface plasmon resonance / mass spectrometry interface for maximum sensitiviityNico Dankbar, Erk Gedig, Simone Königpp 77-80Abstract:An improved interface for coupled protein interaction and detection experiments is presented. A novel flow cell for a surface plasmon resonance (SPR) biosensor carries removable miniaturized sensing pins which can be inserted into MALDI target plates for mass spectrometric (MS) detection of analytes on the sensor surface. Combined with the application of new hydrogel sensor surfaces, the use of the flow cell allows measurements of higher sensitivity and better reproducibility with SPR/MS.Polysiloxane background in field-based ion generation at atmospheric pressureSimone Königpp 81-85Abstract:Polycyclosiloxanes are detected from the laboratory air with field-based ion generation (FBIG) from natural and artificial microemitters (e.g. hairs on fruit flies or sharp metal tips). They produce intense spectra which can be fragmented to generate the same ions which were observed with electron impact mass spectrometry. The abundance of protonated polydimethylcyclosiloxanes is increased under electrospray conditions.

APPLICATION NOTES

Screening methodology for substrates of protein histidine phosphataseKarin Ludwig, Josef Krieglstein, Susanne Klumpp, Simone Königpp 89-98Abstract:Protein histidine phosphatase (PHP) is an enzyme which removes phosphate groups from histidine residues. It was described for vertebrates in the year 2002 and three physiological and one non-physiological protein substrates have been identified so far. Here, a methodology based on affinity purification and mass spectrometry is presented which provided a list of further interaction partners of PHP from rat liver homogenate. Those include hemoglobin subunits and glutathione-S transferase isoforms.Quantification of Imatinib in cell lysatesMarion Viktor, Julio Ciarimboli, Simone Königpp 99-105Abstract:The structure and physico-chemical properties of Imatinib allow its quantification with methodology routinely available in proteomic core facilities. The drug can be separated on C18 material and it is well detectable using UV at 214 nm or mass spectrometry. The paper discusses advantages and disadvantages of either method with respect to drug analysis in cell lysates.

SEQUENCING NOTE

The amino acid sequence of annexin A2 of Syrian hamster Mesocricetus auratus as determined by mass spectrometryUlli-Martin Hohenester, Nina Quiskamp, Ursula Rescher, Simone Königpp 109-117Abstract:The amino acid sequence of annexin A2 from Syrian hamster (Mesocricetus auratus) was investigated by mass spectrometry and aligned with the known annexin A2 sequences available in public databases. The sequence contains specific peptides found in mouse annexin A2 but is mostly homolog to the human protein. It is N-terminally acetylated. At least two phosphorylation sites are indicated.

PROCEEDINGS OF THE 6TH MÜNSTER CONFERENCE ON SINGLE CELL AND MOLECULE ANALYSIS Nov 23-24, 2009

pp 121-138INTRODUCTIONSimone KönigSPEAKER ABSTRACTSThe persistence of memory redoxSteven VogelDevelopments in fluorescence nanoscopyAlexander EgnerCell surface G protein-coupled receptor oligomerization detected using HTRF and Snaptag labeling methodLaurent PrezeauCritical role of dynamic nanodmain organization at the plasma membrane for efficient cell signalingDidier MarguetMultiparameter and imaging flow cytometry: Insights into ummunology and dengue pathogenesisKerstin LühnDynamics of protein-DNA complexes analyzed at the single molecule levelMichael MeisterernstNew molecular tools for specific analysis of nucleic acid and protein biomarkers in single cellsAnders AlderbornProtein microarrays as quantitative tool for the analysis of signal transduction networksUlrike KorfSynthesis of high complexity peptide arrays for proteome researchThomas Felgenhauer, F. R. Bischoff, V. Stadler, F. BreitlingProfiling the parental contribution to the early embryonic transcriptome in ArabidopsisMichael RaissigAnalysis of cellular and molecular interactions using Atomic Force MicroscopyTanja NeumannThe new horizon in proteomics - Introducing high performance electrophoresisReiner WestermeierFrontiers in expression analysis: NuGEN`s ovation RNA amplification at single cell levelWieland KeilholzPOSTER ABSTRACTSClinic InventMarion WillenborgDual-color quantum dot labeling of a heterotetrameric K+ channelDaniel E.J. Waschk, Anke Fabian, Albrecht SchwabNanodiscs in the study of membrane proteinsJulian M. Glück, Marc Wittlich, Sophie Feuerstein, Silke Hoffmann, Dieter Willbold, Bernd W. KönigCytotoxic effects of ergot alkaloids on human primary cellsDennis MUlac, Hans-Ulrich HumpfNMR structure of the "Finger" loop of rod arrestin induced by meta-II rhodopsin bindingSophie Feuerstein, Alexander Pulvermüller, Joachim Granzin, Bernd W. KönigNMR characterization of a transmembrane and cytoplasmic CD4 fragment and its interaction with the soluble domain of HIV-1 VpUMarc Wittlich, Silke Hoffmann, ieter Willbold, Bernd W. KönigDNA replication - A novel function for the B-Myb transcription factorThore Schmedt, Eugen Werwein, Karl-Heinz Klempnauer