Ultrafast Tyrosinase-Mediated Biotinylation of Living Cell Surface Analysis Reveals Novel Cell Surface Proteins Responsible for Influenza A Virus Entry

Cell surface proteins are integral to a myriad of biological
processes,
including cell–cell interactions, signal transduction, and
cell adhesion. Notably, these proteins also serve as key receptors
for numerous pathogens. However, a comprehensive analysis of the surfaceome
remains a significant challenge, primarily due to the high hydrophobicity
and low abundance of these proteins. Here, we developed a novel cell
surface profiling approach, tyrosinase-mediated cell surface labeling
(TYRCSL). Tyrosinase can mildly oxidize a phenol or catechol to the
corresponding o-quinone, which rapidly reacts with
available protein nucleophiles. Leveraging the fast-labeling kinetics
(within 1 min) and minimal toxicity of this method, we explored surfaceome
dynamics during early stages of influenza A virus (IAV) entry. Our
findings reveal that cell surface proteins PODXL2, CNTNAP1, and GPR39
play a crucial role in virus binding and internalization, providing
valuable insights into the molecular mechanisms of IAV entry. TYRCSL
shows the potential to be applied to the system-wide study of the
entry process across a spectrum of pathogens and other instantaneous
surfaceome changes under various stimuli.