Assessing the Multimeric States of Proteins by Matrix-Assisted Laser Desorption Mass Spectrometry: Comparison with Other Biochemical Methods

摘要: Publisher Summary This chapter describes a study conducted to assess the multimeric states of proteins by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). In study, Baker's yeast alcohol dehydrogenase and glucose 6-phosphate were dissolved in water or buffer concentration 10 pmol/μl (10 μM). Glutaraldehyde was added final 0.8%. The reaction mixture incubated at room temperature up 60 min then analyzed MALDI-MS. For other analytical methods, 10-fold molar excess glycine used quench further reactivity glutaraldehyde for mixtures obtain MALDI spectra. UV time-of-flight spectra obtained with Finnigan-MAT Lasermat Vestec VT 2000, both utilizing nitrogen laser. results showed that addition protein is accompanied an increase molecular weight protein. depends upon number reacting groups on side chains accessible cross-linking agent solution intermolecular distance chains.