PrP conformational transitions alter species preference of a PrP-specific antibody

Wen Quan Zou, Jan Langeveld, Xiangzhu Xiao, Shugui Chen, Patrick L. McGeer, Jue Yuan, Michael C. Payne, Hae Eun Kang, John McGeehan, Man Sun Sy, Neil S. Greenspan, David Kaplan, Gong Xian Wang, Piero Parchi, Edward Hoover, Geoff Kneale, Glenn Telling, Witold K. Surewicz, Qingzhong Kong, Jian Ping Guo

Research output: Contribution to journalArticle

Abstract

The epitope of the 3F4 antibody most commonly used in human prion disease diagnosis is believed to consist of residues Met-Lys-His-Met (MKHM) corresponding to human PrP-(109-112). This assumption is based mainly on the observation that 3F4 reacts with human and hamster PrP but not with PrP from mouse, sheep, and cervids, in which Met at residue 112 is replaced by Val. Here we report that, by brain histoblotting, 3F4 did not react with PrP of uninfected transgenic mice expressing elk PrP; however, it did show distinct immunoreactivity in transgenic mice infected with chronic wasting disease. Compared with human PrP, the 3F4 reactivity with the recombinant elk PrP was 2 orders of magnitude weaker, as indicated by both Western blotting and surface plasmon resonance. To investigate the molecular basis of these species- and conformer-dependent preferences of 3F4, the epitope was probed by peptide membrane array and antigen competition experiments. Remarkably, the 3F4 antibody did not react with MKHM but reacted strongly with KTNMK (corresponding to human PrP-(106-110)), a sequence that is also present in cervids, sheep, and cattle. 3F4 also reacted with elk PrP peptides containing KTNMKHV. We concluded that the minimal sequence for the 3F4 epitope consists of residues KTNMK, and the species- and conformer-dependent preferences of 3F4 arise largely from the interactions between Met112 (human PrP) or Val115 (cervid PrP) and adjacent residues.

Original languageEnglish
Pages (from-to)13874-13884
Number of pages11
JournalJournal of Biological Chemistry
Volume285
Issue number18
DOIs
Publication statusPublished - Apr 30 2010

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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    Zou, W. Q., Langeveld, J., Xiao, X., Chen, S., McGeer, P. L., Yuan, J., Payne, M. C., Kang, H. E., McGeehan, J., Sy, M. S., Greenspan, N. S., Kaplan, D., Wang, G. X., Parchi, P., Hoover, E., Kneale, G., Telling, G., Surewicz, W. K., Kong, Q., & Guo, J. P. (2010). PrP conformational transitions alter species preference of a PrP-specific antibody. Journal of Biological Chemistry, 285(18), 13874-13884. https://doi.org/10.1074/jbc.M109.088831