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Mass Spectrometry-Based Methods for Studying Kinetics and Dynamics in Biological Systems

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Book cover Biophysical Techniques in Photosynthesis

Part of the book series: Advances in Photosynthesis and Respiration ((AIPH,volume 26))

In recent years, mass spectrometry (MS) has become one of the most widely used analytical techniques. MS allows studies on compounds ranging in size from single atoms to mega-Dalton biomolecular assemblies. This chapter provides an overview of recent MS applications in biophysical chemistry. The focus of our discussion is on ‘time-resolved’ techniques for tracking changes in complex biological reaction mixtures on time scales of milliseconds to days, thereby providing important structural and mechanistic insights. After a general introduction to biological MS, we discuss practical aspects of time-resolved membrane inlet mass spectrometry (MIMS), such as membrane properties and the use of different sample chambers. The MIMS technique allows online detection of dissolved gases and volatile compounds. It is particularly useful for resolving competing biochemical reactions involving common reactants, because isotopic labeling of substrates can be performed. As examples we present mechanistic studies on Photosystem II, carbonic anhydrase and hydrogenase. In the third part of this chapter we discuss the kinetics and mechanisms of protein folding and unfolding in solution, which can be explored via electrospray ionization mass spectrometry (ESI-MS). On-line coupling of ESI-MS with continuous-flow rapid mixing devices allows monitoring conformational changes of polypeptide chains with millisecond time resolution, as well as the detection and characterization of (un)folding intermediates. Due to its ‘softness’ the ESI process retains even weakly bound noncovalent complexes during the transition into the gas phase, such that protein-protein and protein-ligand interactions can be monitored directly. Additional insights into the conformational dynamics of proteins can be obtained by using time-resolved ESI-MS in conjunction with hydrogen/deuterium exchange methods. It is hoped that this chapter will stimulate the application of time-resolved MS techniques to a wide range of hitherto unexplored research areas.

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Authors and Affiliations

  1. Department of Chemistry, The University of Western Ontario, N6A 5B7, ON, Canada

    Lars Konermann

  2. Max-Planck-Institut für Bioanorganische Chemie, Stiftstraße 34 – 36, D-45470, Mülheim an der Ruhr, Germany

    Johannes Messinger

  3. Research School of Biological Sciences, The Australian National University, ACT 0200, Canberra, Australia

    Warwick Hillier

Authors
  1. Lars Konermann
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  2. Johannes Messinger
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  3. Warwick Hillier
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Editor information

Editors and Affiliations

  1. Leiden University, 9504, 2300 RA, Leiden, Netherlands

    Thijs J. Aartsma  & Jörg Matysik  & 

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Konermann, L., Messinger, J., Hillier, W. (2008). Mass Spectrometry-Based Methods for Studying Kinetics and Dynamics in Biological Systems. In: Aartsma, T.J., Matysik, J. (eds) Biophysical Techniques in Photosynthesis. Advances in Photosynthesis and Respiration, vol 26. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-8250-4_9

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