TY - JOUR
T1 - Identification and Characterization of Complex Glycosylated Peptides Presented by the MHC Class II Processing Pathway in Melanoma
AU - Malaker, Stacy A.
AU - Ferracane, Michael J.
AU - Depontieu, Florence R.
AU - Zarling, Angela L.
AU - Shabanowitz, Jeffrey
AU - Bai, Dina L.
AU - Topalian, Suzanne L.
AU - Engelhard, Victor H.
AU - Hunt, Donald F.
N1 - Funding Information:
We wish to acknowledge Tracee L. McMiller and Theresa S. Pritchard (Johns Hopkins University School of Medicine) for technical assistance. Additionally, we would like to thank Jane V. Aldrich (University of Florida Department of Medicinal Chemistry) for providing access to Molecular Operating Environment (MOE). This work was supported by a research grant from the Melanoma Research Alliance (to S.L.E, D.F.H. and V.H.E) and by National Institutes of Health Grant AI033993 (to D.F.H.). This work was also supported by USPHS Grants R01 A120963 and CA134060 (to V.H.E.). A.L.Z. was a recipient of a Sidney Kimmel Scholar award.
Publisher Copyright:
© 2016 American Chemical Society.
PY - 2017/1/6
Y1 - 2017/1/6
N2 - The MHC class II (MHCII) processing pathway presents peptides derived from exogenous or membrane-bound proteins to CD4+ T cells. Several studies have shown that glycopeptides are necessary to modulate CD4+ T cell recognition, though glycopeptide structures in these cases are generally unknown. Here, we present a total of 93 glycopeptides from three melanoma cell lines and one matched EBV-transformed line with most found only in the melanoma cell lines. The glycosylation we detected was diverse and comprised 17 different glycoforms. We then used molecular modeling to demonstrate that complex glycopeptides are capable of binding the MHC and may interact with complementarity determining regions. Finally, we present the first evidence of disulfide-bonded peptides presented by MHCII. This is the first large scale study to sequence glyco- and disulfide bonded MHCII peptides from the surface of cancer cells and could represent a novel avenue of tumor activation and/or immunoevasion.
AB - The MHC class II (MHCII) processing pathway presents peptides derived from exogenous or membrane-bound proteins to CD4+ T cells. Several studies have shown that glycopeptides are necessary to modulate CD4+ T cell recognition, though glycopeptide structures in these cases are generally unknown. Here, we present a total of 93 glycopeptides from three melanoma cell lines and one matched EBV-transformed line with most found only in the melanoma cell lines. The glycosylation we detected was diverse and comprised 17 different glycoforms. We then used molecular modeling to demonstrate that complex glycopeptides are capable of binding the MHC and may interact with complementarity determining regions. Finally, we present the first evidence of disulfide-bonded peptides presented by MHCII. This is the first large scale study to sequence glyco- and disulfide bonded MHCII peptides from the surface of cancer cells and could represent a novel avenue of tumor activation and/or immunoevasion.
KW - MHC class II
KW - glycopeptide analysis
KW - immunology
KW - mass spectrometry
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U2 - 10.1021/acs.jproteome.6b00496
DO - 10.1021/acs.jproteome.6b00496
M3 - Article
C2 - 27550523
AN - SCOPUS:85017580347
SN - 1535-3893
VL - 16
SP - 228
EP - 237
JO - Journal of proteome research
JF - Journal of proteome research
IS - 1
ER -