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.