title: Electrospray ionization-mass spectrometry characterization of heterotetrameric sarcosine oxidase.
abstract: Electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry has been used to characterize heterotetrameric corynebacterial sarcosine oxidase. By using a conventional quadrupole mass spectrometer, no spectra for the intact complex could be obtained (i.e., electrospraying protein at neutral pH), but spectra showing the four protein subunits were obtained when electrospraying from acidic solution. Initial low resolution ESI-FTICR mass spectra of the intact heterotetramer revealed a typical narrow charge state distribution in the range 6000 < m/z < 9000, consistent with retention of a compact structure in the gas phase, and gave a mass measurement about 1000 u higher than predicted. Efficient in-trap clean up, based upon low energy collisionally induced dissociation of adducts, allowed significant improvement in mass measurement accuracy. The present results represent the largest heteromultimeric protein complex successfully analyzed using FTICR mass spectrometry, and clearly illustrate the importance of sample clean up methods for large molecule characterization.
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
authors.forename: L
authors.initials: L
authors.lastname: Pasa Tolić
authors.forename: A C
authors.initials: AC
authors.lastname: Harms
authors.forename: G A
authors.initials: GA
authors.lastname: Anderson
authors.forename: R D
authors.initials: RD
authors.lastname: Smith
authors.forename: A
authors.initials: A
authors.lastname: Willie
authors.forename: M S
authors.initials: MS
authors.lastname: Jorns
mesh_terms: D003352:Corynebacterium / Q000201:enzymology; D007527:Isoenzymes / Q000032:analysis*; D013058:Mass Spectrometry; D010089:Oxidoreductases, N-Demethylating / Q000032:analysis*; D050879:Sarcosine Oxidase
chemical_list: D007527:Isoenzymes; D010089:Oxidoreductases, N-Demethylating; D050879:Sarcosine Oxidase