Characterization of PA–protein
conjugates formed by NCL.
(a,b) SDS-PAGE visualized by silver staining (a) and Western blot
against GFP (b) were used to analyze the NCL reaction between YFP
and PA (Lane 1: YFP only, Lane 2: YFP conjugated to CysPA, Lane 3:
CysPA only, Lanes L1, L2, L3: protein standards with indicated molecular
weights in kDa). (c) YFP fluorescence intensity after conjugation
to CysPA-coated surface (CysPA-YFP NCL). An identical PA lacking cysteine
(GlyPA-YFP Control) or CysPA treated with reaction buffer alone (CysPA
Control) was used as a control (** p < 0.01 vs
CysPA-YFP NCL). (d) Confocal fluorescence microscopy of CysPA nanofiber-coated
alginate microparticles simultaneously reacted with YFP and CFP. (e)
Fluorescence anisotropy of YFP following NCL reaction with CysPA.
GlyPA was used as a control (* p < 0.05 vs YFP; #p < 0.05 vs GlyPA).