naa a22 4500 510738478 CHVBK 20180411083019.0 cr unu---uuuuu 180411e20130601xx s 000 0 eng 10.1007/s12127-013-0126-4 doi (NATIONALLICENCE)springer-10.1007/s12127-013-0126-4 Ligand binding and unfolding of tryptophan synthase revealed by ion mobility-tandem mass spectrometry employing collision and surface induced dissociation [Elektronische Daten] [Royston Quintyn, Mowei Zhou, Shai Dagan, John Finke, Vicki Wysocki] Understanding protein tertiary and quaternary structures is crucial to a better understanding of their biological functions. Here we illustrate for tryptophan synthase that tandem mass spectrometry (MS/MS) reveals not only protein subunit architectures, but also protein unfolding behavior when coupled with ion mobility (IM). In the present study, we verified the subunit arrangement with surface induced dissociation (SID). We are able to correlate experimental results by IM with those obtained in unfolding simulations for the hetero-tetramer Tryptophan Synthase (TS) protein complex by identifying the presence of at least three stable intermediates (I1, I2, and I3) during the unfolding process in collision induced dissociation (CID). We illustrate that the unfolding of the TS complex is likely due to the initial unfolding of an α-monomer subunit, followed by the unfolding of the second α-monomers. We also illustrate the ability of this combination of techniques to not only identify conformational changes of TS upon addition of D,L-α-glycerol phosphate (GP), but also to determine the location of the ligand, which is buried within the α-monomer of the TS. Springer-Verlag Berlin Heidelberg, 2013 Trytophan synthase nationallicence Ligand binding nationallicence Protein unfolding nationallicence Conformational change nationallicence Surface induced dissociation nationallicence Ion mobility nationallicence Quintyn Royston Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut Zhou Mowei Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut Dagan Shai Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut Finke John Department of Chemistry, Oakland University, 260 Science and Engineering Building, Rochester, MI, USA aut Wysocki Vicki Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut International Journal for Ion Mobility Spectrometry Springer-Verlag 16/2(2013-06-01), 133-143 1435-6163 16:2<133 2013 16 12127 https://doi.org/10.1007/s12127-013-0126-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12127-013-0126-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Quintyn Royston Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut NATIONALLICENCE 700 1- Zhou Mowei Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut NATIONALLICENCE 700 1- Dagan Shai Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut NATIONALLICENCE 700 1- Finke John Department of Chemistry, Oakland University, 260 Science and Engineering Building, Rochester, MI, USA aut NATIONALLICENCE 700 1- Wysocki Vicki Department of Chemistry and Biochemistry, University of Arizona, 1306 E. University Blvd., PO Box 210041, Tucson, AZ, USA aut NATIONALLICENCE 773 0- International Journal for Ion Mobility Spectrometry Springer-Verlag 16/2(2013-06-01), 133-143 1435-6163 16:2<133 2013 16 12127 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer