Mass spectrometric analysis of cyanogen bromide fragments of integral membrane proteins at the picomole level: application to rhodopsin

Analytical Biochemistry
P KraftE A Dratz

Abstract

Advances in time-of-flight mass spectrometry allow unit mass resolution of proteins and peptides up to about 6000 Da molecular weight. Identification of larger proteins and study of their posttranslational or experimental modifications by mass analysis is greatly enhanced by cleavage into smaller fragments. Most membrane proteins are difficult to mass analyze because of their high hydrophobicity, typical expression in low quantities, and because the detergents commonly used for solubilization may be deleterious to mass analysis. Cleavage with cyanogen bromide is beneficial for analysis of membrane proteins since the methionine cleavage sites are typically located in hydrophobic domains and cleavage at these points reduces the size of the hydrophobic fragments. Cyanogen bromide also gives high cleavage yields and introduces only volatile contaminants. Even after cleavage membrane proteins often contain fragments that are difficult to chromatograph. Matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) is capable of analyzing complex mixtures without chromatography. We present a MALDI MS method that quickly and reliably identifies the cyanogen bromide fragments and posttranslational modifications of reduced and...Continue Reading

References

Jan 1, 1985·Annual Review of Biochemistry·R Kornfeld, S Kornfeld
Oct 23, 1973·Biochemistry·K Hong, W L Hubbell
May 21, 1974·Biochemistry·D S Papermaster, W J Dreyer
Sep 1, 1995·Biophysical Chemistry·K NakanishiM F Boehm
Apr 16, 1998·Protein Science : a Publication of the Protein Society·L E BallD R Knapp
Apr 12, 2000·Biochemistry·J le CoutreK F Faull
Jun 24, 2000·Science·B BorhanK Nakanishi
Aug 5, 2000·Science·K PalczewskiM Miyano

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Citations

Jan 20, 2009·Biochemistry·George K E UmanahJeffrey M Becker
Jul 20, 2007·Analytical and Bioanalytical Chemistry·Bernhard GranvoglLutz Andreas Eichacker
May 30, 2006·Protein Science : a Publication of the Protein Society·Richard B JacobsenJoseph S Schoeniger
Oct 17, 2009·Proteomics·Ingrid MillerElisabetta Gianazza
Jul 29, 2008·Brain Research·Kenneth M WannemacherRichard D Howells
Apr 13, 2002·Photochemistry and Photobiology·Zsolt AblonczyRosalie K Crouch
Mar 31, 2004·Rapid Communications in Mass Spectrometry : RCM·Ying-Qing YuJohn C Gebler
Aug 28, 2001·Journal of Mass Spectrometry : JMS
Jan 29, 2008·Nature Structural & Molecular Biology·Markus A SeegerKlaas M Pos
Oct 4, 2006·The Journal of Biological Chemistry·Ross M TaylorAlgirdas J Jesaitis
Dec 25, 2004·Proteomics·Christian KleinDieter Oesterhelt
Apr 29, 2003·Journal of Proteome Research·Michael B GosheRichard D Smith
May 3, 2007·Journal of Proteome Research·Nikolai ZvonokAlexandros Makriyannis

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