Observation of noncovalent complexes using laser-induced liquid beam ionization/desorption

Kleinekofort, Wolfgang; Pfenninger, Anja; Plomer, Thomas; Griesinger, Christian; Brutschy, Bernhard

doi:10.1016/s0168-1176(96)04507-7

Cited by 48 publications

(33 citation statements)

References 16 publications

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“…Another important aspect is its tolerance to various buffers in solution, which often play an important role in determining binding selectivity and strength of DNA/RNA-ligand complexes. LILBID in its present form is a development from its earlier version [25][26][27][28] using liquid beams, which suffered from a very poor duty cycle.…”

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confidence: 99%

Binding sites of the viral RNA element TAR and of TAR mutants for various peptide ligands, probed with LILBID: A new laser mass spectrometry

Morgner

Barth

Brutschy

et al. 2008

J. Am. Soc. Mass Spectrom.

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A new laser-based mass spectrometry method, called laser induced liquid bead ion desorption (LILBID), was applied to investigate RNA:ligand interactions. As model system the HIV Tat:TAR transactivation complex and its binding behavior were analyzed. TARwt of HIV Type 1 and Type 2 and different artificial mutants were compared regarding their binding to Tat and different peptide ligands. Specific and nonspecific association to TAR was deduced, with the bulge being the well known specific binding site of TAR. In the case of triple arginine (RRR) as a nonspecific ligand, multiple electrostatic binding to TAR was found at higher concentration of RRR. This contrasted with the formation of only ternary complexes in competitive binding studies with TAR, Tat, and potential inhibitors. The fact that the stoichiometries of the complexes can be determined is a major advantage of MS methods over the widely applied fluorimetric methods. A quantitative evaluation of the spectra by a numeric model for ternary complex formation allows conclusions about the role and strength of the binding sites of the RNAs, the specificity and affinity of different ligands, the determination of apparent IC50 and KD values, and a comparison of the binding efficiencies of potential inhibitors.

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confidence: 99%

Binding sites of the viral RNA element TAR and of TAR mutants for various peptide ligands, probed with LILBID: A new laser mass spectrometry

Morgner

Barth

Brutschy

et al. 2008

J. Am. Soc. Mass Spectrom.

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“…A classic solid matrix was still employed in the first IR-MALDI-MS experiments by Hillenkamp and coworkers [8] for the purpose of compatibility with the vacuum of the mass spectrometer, whereas in the Brutschy group, a liquid water/alcohol mixture was directly introduced into the high vacuum chamber of a time-of-flight (ToF) mass spectrometer in the form of a microbeam. The ions were released by IR laser excitation of an alcoholic C-O vibration (λ = 10.6 μm, CO 2 -Laser) [9,10]. It was later possible to analyze pure water solutions by utilizing infrared radiation of an Er:YAG laser or an optical parametric oscillator (OPO) to directly excite the O-H stretch vibration mode [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

IR-MALDI ion mobility spectrometry

et al. 2016

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The novel combination of infrared matrix-assisted laser dispersion and ionization (IR-MALDI) with ion mobility (IM) spectrometry makes it possible to investigate biomolecules in their natural environment, liquid water. As an alternative to an ESI source, the IR-MALDI source was implemented in an in-house-developed ion mobility (IM) spectrometer. The release of ions directly from an aqueous solution is based on a phase explosion, induced by the absorption of an IR laser pulse (λ = 2.94 μm, 6 ns pulse width), which disperses the liquid as nano- and micro-droplets. The prerequisites for the application of IR-MALDI-IM spectrometry as an analytical method are narrow analyte ion signal peaks for a high spectrometer resolution. This can only be achieved by improving the desolvation of ions. One way to full desolvation is to give the cluster ions sufficient time to desolvate. Two methods for achieving this are studied: the implementation of an additional drift tube, as in ESI-IM-spectrometry, and the delayed extraction of the ions. As a result of this optimization procedure, limits of detection between 5 nM and 2.5 μM as well as linear dynamic ranges of 2-3 orders of magnitude were obtained for a number of substances. The ability of this method to analyze simple mixtures is illustrated by the separation of two different surfactant mixtures.

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“…In a series of publications Brutschy and co-workers have shown that this method has a great potential for the analysis of macro-and biomolecules and their complexes even in the liquid phase, 1,7,[11][12][13][14][15][16][17] a feature that is less obvious and not straightforward for the standard and nowadays wellestablished techniques like electrospray ionization 18 or matrix-assisted laser desorption/ionization ͑MALDI͒ mass spectrometry. 19 In this article we describe a new LILBID-tandem mass spectrometer ͑MS͒ apparatus whose mass resolution has been increased significantly.…”

Section: Introductionmentioning

confidence: 99%

New design for a time-of-flight mass spectrometer with a liquid beam laser desorption ion source for the analysis of biomolecules

Charvát

Lugovoj

Faubel³

et al. 2004

Review of Scientific Instruments

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Articles you may be interested inIon spectrometer composed of time-of-flight and Thomson parabola spectrometers for simultaneous characterization of laser-driven ions Rev. Sci. Instrum. 80, 053302 (2009); 10.1063/1.3131628 High-energy collision induced dissociation fragmentation pathways of peptides, probed using a multiturn tandem time-of-flight mass spectrometer "MULTUM-TOF/TOF" Rev. Sci. Instrum. 78, 074101 (2007); 10.1063/1.2751403 Simple and inexpensive time-of-flight charge-to-mass analyzer for ion beam source characterization Rev. Sci. Instrum. 77, 063301 (2006);We describe a novel liquid beam mass spectrometer, based on a recently discovered nanosecond laser desorption phenomenon, ͓W. Kleinekofort, J. Avdiev, and B. Brutschy, Int. J. Mass Ion. Processes 152, 135 ͑1996͔͒ which allows the liquid-to-vacuum transfer, and subsequent mass analysis of pre-existing ions and ionic associates from liquid microjets of aqueous solutions. The goal of our novel technical approach is to establish a system with good mass resolution that implements improvements on critical components that make the system more reliable and easier to operate. For laser desorption pulsed dye-laser difference frequency mixing is used that provides tunable infrared light near the absorption maximum of liquid water around 3 m. Different types of liquid beam glass nozzles ͑convergent capillary and aperture plate nozzles͒ are investigated and characterized. Starting from theoretical considerations of hydrodynamic drag forces on micrometer size droplets in supersonic rarefied gas flows we succeeded in capturing efficiently the liquid beam in a liquid beam recycling trap operating at the vapor pressure of liquid water. For improving the pollution resistance, the liquid jet high vacuum ion source region is spatially separated from the reflectron time-of-flight mass spectrometer ͑TOF-MS͒ working behind a gate valve in an ultrahigh vacuum environment. A simple ͑simulation optimized͒ ion optics is employed for the ion transfer from the source to the high vacuum region. This new feature is also mostly responsible for the improved mass resolution. With the present tandem-TOF-MS setup a resolution of m/⌬mϷ1800 for the low and m/⌬mϷ700 in the high mass region has been obtained for several biomolecules of different mass and complexity ͑amino acids, insulin, and cytochrome c͒.

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Observation of noncovalent complexes using laser-induced liquid beam ionization/desorption

Cited by 48 publications

References 16 publications

Binding sites of the viral RNA element TAR and of TAR mutants for various peptide ligands, probed with LILBID: A new laser mass spectrometry

Binding sites of the viral RNA element TAR and of TAR mutants for various peptide ligands, probed with LILBID: A new laser mass spectrometry

IR-MALDI ion mobility spectrometry

New design for a time-of-flight mass spectrometer with a liquid beam laser desorption ion source for the analysis of biomolecules

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