Figure 4

(A) ELISA showing that both wTMV (blue) and mTMV (red) have similar binding to polyclonal TMV antibodies (CAB 57400/1000). The correlation coefficient between the two curves is 0.998 using Pearson correlation coefficient, suggesting that there is no difference in antibody binding. The error bars show ± 2 standard deviations with a sample size n = 3. (B) Schematic showing the conjugation of diazonium salt with alkyne handle via diazonium coupling on the Y139. Conjugation should increase coat protein mass by 128 Da. This figure was made using Chemdraw 20.0 (https://perkinelmerinformatics.com/products/research/chemdraw/). (C) Deconvoluted ESI–MS spectra of TMV-Alkyne from both predominantly wTMV and mTMV. The wTMV-Alk shows a peak at 17,662 Da and a preceding small peak at 17,645 Da. The mTMV-Alk generates four peaks at 17,645 Da, 17,662 Da, 17,675 Da, and 17,692 Da. The first two peaks from the left, 17,645 Da and 17,662 Da, belong to wTMV-Alk. The last two peaks, 17,675 Da and 17,692 Da, belong to mTMV-Alk. (D) TEM showing that the morphology of TMV rods with alkyne remain intact. These results suggest that tyrosine functionalization is unaffected by this mutation.