Selective Enrichment of Histidine Phosphorylated Peptides Using Molecularly Imprinted Polymers.
Anal Chem
; 93(8): 3857-3866, 2021 03 02.
Article
en En
| MEDLINE
| ID: mdl-33591162
ABSTRACT
Protein histidine phosphorylation (pHis) is involved in molecular signaling networks in bacteria, fungi, plants, and higher eukaryotes including mammals and is implicated in human diseases such as cancer. Detailed investigations of the pHis modification are hampered due to its acid-labile nature and consequent lack of tools to study this post-translational modification (PTM). We here demonstrate three molecularly imprinted polymer (MIP)-based reagents, MIP1-MIP3, for enrichment of pHis peptides and subsequent characterization by chromatography and mass spectrometry (LC-MS). The combination of MIP1 and ß-elimination provided some selectivity for improved detection of pHis peptides. MIP2 was amenable to larger pHis peptides, although with poor selectivity. Microsphere-based MIP3 exhibited improved selectivity and was amenable to enrichment and detection by LC-MS of pHis peptides in tryptic digests of protein mixtures. These MIP protocols do not involve any acidic solvents during sample preparation and enrichment, thus preserving the pHis modification. The presented proof-of-concept results will lead to new protocols for highly selective enrichment of labile protein phosphorylations using molecularly imprinted materials.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Impresión Molecular
/
Histidina
Idioma:
En
Revista:
Anal Chem
Año:
2021
Tipo del documento:
Article