Electrospray ionization mass spectrometry of highly heterogeneous protein systems: protein ion charge state assignment via incomplete charge reduction.
نویسندگان
چکیده
Correct mass and charge assignment for large highly heterogeneous macromolecular ions (e.g., large glycoproteins with significant carbohydrate content) presents a great challenge in native electrospray ionization mass spectrometry (ESI MS). A new approach to this problem combines complexity reduction (mass-selection of a narrow distribution of ionic species from a heterogeneous mixture) and gas-phase ion chemistry (electron-transfer reactions) to induce partial reduction of the ionic charge. The resulting spectra are devoid of complexity and are easy to interpret, leading to correct mass assignment. The new method is tested using several glycoproteins and their complexes, for which standard deconvolution approaches do not work.
منابع مشابه
Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes.
Native electrospray-ionization mass spectrometry (native MS) measures biomolecules under conditions that preserve most aspects of protein tertiary and quaternary structure, enabling direct characterization of large intact protein assemblies. However, native spectra derived from these assemblies are often partially obscured by low signal-to-noise as well as broad peak shapes because of residual ...
متن کاملEnhancing Accuracy in Molecular Weight Determination of Highly Heterogeneously Glycosylated Proteins by Native Tandem Mass Spectrometry
The determination of molecular weights (MWs) of heavily glycosylated proteins is seriously hampered by the physicochemical characteristics and heterogeneity of the attached carbohydrates. Glycosylation impacts protein migration during sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and size-exclusion chromatography (SEC) analysis. Standard electrospray ionization (ESI)-ma...
متن کاملA quadrupole ion trap mass spectrometer with three independent ion sources for the study of gas-phase ion/ion reactions.
An instrument for the study of gas-phase ion/ion reactions in which three independent sources of ions, namely, two electrospray ionization sources and one atmospheric sampling glow discharge ionization source, are interfaced to a quadrupole ion trap mass analyzer is described. This instrument expands the scope of gas-phase ion/ion reaction studies by allowing for manipulation of the charge stat...
متن کاملEffects of anions on the positive ion electrospray ionization mass spectra of peptides and proteins.
Positive ion electrospray ionization mass spectra of polypeptides are usually obtained from solutions that are acidified and therefore contain relatively high concentrations of anions. The present study describes an investigation of the effects of these ubiquitous anions on the positive ion electrospray ionization mass spectra of peptides and proteins. Certain anionic species in the spray solut...
متن کاملEffect of protein stabilization on charge state distribution in positive- and negative-ion electrospray ionization mass spectra.
Changes in protein conformation are thought to alter charge state distributions observed in electrospray ionization mass spectra (ESI-MS) of proteins. In most cases, this has been demonstrated by unfolding proteins through acidification of the solution. This methodology changes the properties of the solvent so that changes in the ESI-MS charge envelopes from conformational changes are difficult...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Analytical chemistry
دوره 82 18 شماره
صفحات -
تاریخ انتشار 2010