Selective enrichment and analysis of acidic peptides and proteins using polymeric reverse micelles and MALDI-MS.
The typical difficulties associated with the detection of acidic peptides (i.e., those with low isoelectric points (pI)) by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) represent a challenge in some proteomic analyses. Here, reverse micelle-forming amphiphilic homopolymers with positively charged interiors are synthesized and used to selectively enrich low pI peptides from complex mixtures for MALDI-MS detection. When using these polymers, acidic proteolytic peptides that are undetectable during normal MALDI-MS analysis are selectively detected. We show that enrichment of these low pI peptides allows acidic proteins to be selectively targeted for detection in multiprotein digests. In addition, the presence of the positively charged polymers during MALDI-MS analyses enhances peptide ion signals by almost an order of magnitude, thereby achieving reproducible ion signals for acidic peptides at concentrations as low as 100 fM. Concurrent detection of acidic and basic peptides was also facilitated by utilizing a sequential extraction process involving reverse micelle forming polymers with positively and negatively charged interiors.
Rodthongkum
Nadnudda
N
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Chen
Yangbin
Y
Thayumanavan
S
S
Vachet
Richard W
RW
D000143:Acids / Q000737:chemistry; D000818:Animals; D002417:Cattle; D002645:Chickens; D006736:Horses; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008823:Micelles; D015394:Molecular Structure; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000143:Acids; D008823:Micelles; D010455:Peptides; D011506:Proteins