Fig 3.

Epitope mapping on a microarray of the immunodominant sequence in the region of amino acids 491 to 510. (A) Glycopeptide walk of GalNAc attachment sites, including scan peptide 1 (491-510), four single-site synthetic analogues (2 to 5) (glycosylation sites are underlined and in boldface), and glycopeptide 6, produced by GalNAc-T2 glycosylation of glycopeptide 1 in solution (potential glycosylation sites are underlined and italicized). MALDI-TOF analysis confirmed five out of six possible glycosylation sites. (B) Alanine mutation walk within the region of amino acids 500 to 510 (the alanine mutation site is italicized, and the glycosylation sites are underlined and in boldface). (C) Single-amino-acid epitope walk over the region of amino acids 484 to 524 with fixed glycosylation at Thr⁵⁰⁴ (glycosylation sites are underlined and in boldface). (D) Analysis of different glycoisoforms with focused 7-mer peptide ⁵⁰¹PPAXAPG⁵⁰⁷ analogues (T/S = Ser or Thr). Microarray data presented for a representative patient serum (number 63; HSV-1/-2 positive) at a 1:60 dilution. Each value is the average of four replicates, with the standard deviation shown by error bars. See also Fig. S2 and S3 in the supplemental material for the full data set.