Dear all,
I am conducting research on site-directed mutagenesis of crystalline proteins. My aim is to verify the desired mutation in the protein sequence using the MALDI-TOF mass spectrometry technique. I would like to know if this method is feasible for my research.
Thank you.
With a purified, intact protein of uniform size, the resolution of basic MALDI-TOF should be sufficient to tell whether the molecular weight of the protein has changed by the expected amount, give or take a few daltons, as long as the protein is not too large.
Just wanted to know what kind of mutation you want to detect with MALDI-TOF? As mutating Leu to Iso will not change the molecular weight of protein, and similarly, Arg and Asp has a unit dalton difference.
@ Adam B Shapiro , I appreciate your great guidance. When analyzing the acquired mass spectrum data, I found an m/z value that may identify the mutation in the raw data, located near the main peak. What do you think, is this a valid approach to data analysis?
Thank you for your answer. Also , I have been unable to find a paper related to the application of MALDI-TOF mass spectrometry for characterizing single amino acid variants of proteins. Can you guide me on this?" regards @Ayaz Anwar
Farid Nasiri I think it is a valid analysis, but you may want to do a more detailed investigation. For example, you can digest the protein with proteases and study the masses of the resulting peptides.
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