NR AJSL
AU Prusiner,S.B.; Groth,D.F.; Bolton,D.C.; Kent,S.B.; Hood,L.E.
TI Purification and structural studies of a major scrapie prion protein
QU Cell 1984 Aug; 38(1): 127-34
PT journal article
AB Scrapie is a degenerative, neurological disorder caused by a slow infectious agent or prion. Extensively purified preparations of prions were denatured by boiling in sodium dodecyl sulfate and the major protein component (PrP 27-30) was isolated by preparative HPLC size exclusion chromatography after proteinase K digestion. The purified PrP 27-30 molecules were not infectious. Ultraviolet absorption spectra of purified PrP 27-30 demonstrated the absence of covalently linked polynucleotides. Amino acid composition studies showed that PrP 27-30 contains at least 17 naturally occurring amino acids. A single N-terminal amino acid sequence for PrP 27-30 was obtained; the sequence is N-Gly-Gln-Gly-Gly-Gly-Thr-His-Asn-Gln-Trp-Asn-Lys-Pro-Ser-Lys and it does not share homology with any known proteins. The same amino acid sequence was found when an extensively purified preparation of prions aggregated into rods and containing approximately 10(9.5) ID50 U/ml was sequenced directly. Knowledge of the amino acid sequence should permit determination of the genetic origin and replication mechanism of prions.
IN Intensiv gereinigte Prion-Präparationen wurden durch Kochen in SDS denaturiert und mit Proteinase K verdaut. Dadurch wurden aus den Prionen Prionproteine mit scheinbaren Molekularmassen von 27-30 kDa isoliert, welche nicht infektiös waren. Das UV-Licht-Absorptionsspektrum der gereinigten PrP27-30 sprach gegen die Anwesenheit von Nukleinsäuren. Die aminoterminale Sequenz des PrP27-30 war N-Gly-Gln-Gly-Gly-Gly-Thr-His-Asn-Gln-Trp-Asn-Lys-Pro-Ser-Lys.
MH Amino Acid Sequence; Amino Acids/analysis; Animal; Brain/microbiology; Electrophoresis, Polyacrylamide Gel; Hamsters; Molecular Weight; Prions/*analysis; Spectrophotometry, Ultraviolet; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; Viral Proteins/*isolation & purification
AD Stanley B. Prusiner, Department of Neurology and Biochemistry and Biophysics, University of California, San Francisco 94143, California
SP englisch
PO USA
OR Prion-Krankheiten 6