TY - JOUR
T1 - Thr but Asn of the N-glycosylation sites of PrP is indispensable for its misfolding
AU - Ikeda, Shino
AU - Kobayashi, Atsushi
AU - Kitamoto, Tetsuyuki
N1 - Funding Information:
This study was supported by the Program for Promotion of Fundamental Studies in Health Sciences of National Institute of Biomedical Innovation (T.K.), a grant from the Ministry of Health, Labor and Welfare (A.K. and T.K.), and a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (A.K. and T.K.). In addition, we thank R.-W. Shin, M. Morita and B. Bell for critical review of the manuscript.
PY - 2008/5/16
Y1 - 2008/5/16
N2 - Prion protein (PrP) contains two N-linked glycosylation sites. It is unknown which amino acid substitution contributes most efficiently to the abolishment of N-linked glycosylations. To define the influence of amino acid substitution at the N-linked glycosylation sites on the conversion efficiency of mouse PrP, we tested each of all 19 amino acid substitutions at either one of the N-linked glycosylation sites (codon 180, 182, 196 or 198). The conversion efficiency of the mutagenized PrP was highly dependent on the newly introduced amino acid itself regardless of the absence of N-linked glycosylation in scrapie-infected mouse neuroblastoma cells. The majority of mutant PrP with substitutions at the Asn residues of the N-linked glycosylation sites were conversion-competent, whereas most mutant PrP with substitutions at the Thr residues were conversion-incompetent. These findings emphasize that the Asn residues of the N-linked glycosylation sites are replaceable to abolish N-linked glycosylations without directly affecting the protein function.
AB - Prion protein (PrP) contains two N-linked glycosylation sites. It is unknown which amino acid substitution contributes most efficiently to the abolishment of N-linked glycosylations. To define the influence of amino acid substitution at the N-linked glycosylation sites on the conversion efficiency of mouse PrP, we tested each of all 19 amino acid substitutions at either one of the N-linked glycosylation sites (codon 180, 182, 196 or 198). The conversion efficiency of the mutagenized PrP was highly dependent on the newly introduced amino acid itself regardless of the absence of N-linked glycosylation in scrapie-infected mouse neuroblastoma cells. The majority of mutant PrP with substitutions at the Asn residues of the N-linked glycosylation sites were conversion-competent, whereas most mutant PrP with substitutions at the Thr residues were conversion-incompetent. These findings emphasize that the Asn residues of the N-linked glycosylation sites are replaceable to abolish N-linked glycosylations without directly affecting the protein function.
KW - Conversion
KW - N-linked glycosylation
KW - Prion protein
KW - Site-directed mutagenesis
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U2 - 10.1016/j.bbrc.2008.03.014
DO - 10.1016/j.bbrc.2008.03.014
M3 - Article
C2 - 18343219
AN - SCOPUS:41149163562
VL - 369
SP - 1195
EP - 1198
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 4
ER -