Light, pH, and Temperature Triple-Responsive Magnetic Composites for Highly Efficient Phosphopeptide Enrichment

Abstract

Smart materials can dynamically and reversibly change their structures and functions in response to external stimuli. In this study, we designed a smart magnetic composite (MNP-pSPA-b-pNIPAm) with a triple response to ultraviolet (UV) light, pH, and temperature. Relying on the response of spiropyranyl acrylate (SPA) and N-isopropylacrylamide (NIPAm) to external stimuli (light, pH, and temperature), MNP-pSPA-b-pNIPAm was used for the controlled capture and release of phosphopeptides. The established phosphopeptide enrichment platform exhibits high sensitivity (detection limit of 0.04 fmol), high selectivity (BSA/β-casein, 1000:1), and good reusability (6 cycles). In addition, the method was also applied to the enrichment of phosphopeptides in real samples (skim milk, human saliva, and serum), demonstrating the feasibility of this method for phosphoproteomic analysis. After enriching from human nonsmall cell lung cancer cell (A549) lysates with MNP-pSPA-b-pNIPAm, 2595 phosphopeptides corresponding to 2281 phosphoproteins were identified. The novel responsive enrichment probe is highly specific for phosphoproteomic analysis and provides an effective method for studying the significance of protein phosphorylation in complex biological samples.

Innovative Chemical Tools to Address Analytical Challenges of Protein Phosphorylation and Glycosylation

“…Similar to our strategy, Chen et al reported a 2D supramolecular polymer with a controlled disassembly and reassembly property for efficient capture and release of PPs . Besides, inspired by the design strategy of this kind of smart polymers, the groups of Zhu and Jia further advanced the development of phosphorylation enrichment materials by combining a smart polymer with magnetic beads or introducing a new responsive group (e.g., spiropyran). , …”

Enrichment of Phosphopeptides Based on Zirconium Phthalocyanine-Modified Magnetic Nanoparticles

“…Post-translational modifications (PTMs) of proteins greatly increase the complexity of gene products and play an important role in multiple cellular processes. , Protein phosphorylation, as one of the most essential of PTMs, performs important functions in numerous biological events. , Protein phosphorylation is a highly complex and dynamic process. Abnormal phosphorylation regulation is related to a variety of human diseases, such as leukemia, neurodegenerative diseases, etc. , As a result, the study of protein phosphorylation is vital for identifying cancer markers and discovering drug targets.…”

Cotton Ti-IMAC: Developing Phosphorylated Cotton as a Novel Platform for Phosphopeptide Enrichment

“…Various techniques have been established for phosphopeptide enrichment so far. , Among them, immobilized metal affinity chromatography (IMAC) has become the most competent and widely used one. IMAC was first developed in 1975 by Porath et al to fractionate protein solutions based on the transition metal ion affinity .…”