Prusiner S B, Groth D F, Bolton D C, Kent S B, Hood L E
Cell. 1984 Aug;38(1):127-34. doi: 10.1016/0092-8674(84)90533-6.
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.
羊瘙痒症是一种由缓慢感染因子或朊病毒引起的退行性神经疾病。将朊病毒的高度纯化制剂在十二烷基硫酸钠中煮沸使其变性，蛋白酶K消化后，通过制备型高效液相色谱尺寸排阻色谱法分离出主要蛋白质成分（PrP 27-30）。纯化后的PrP 27-30分子没有传染性。纯化后的PrP 27-30的紫外吸收光谱表明不存在共价连接的多核苷酸。氨基酸组成研究表明，PrP 27-30含有至少17种天然存在的氨基酸。获得了PrP 27-30的单一N端氨基酸序列；该序列为N-甘氨酸-谷氨酰胺-甘氨酸-甘氨酸-甘氨酸-苏氨酸-组氨酸-天冬酰胺-谷氨酰胺-色氨酸-天冬酰胺-赖氨酸-脯氨酸-丝氨酸-赖氨酸，且与任何已知蛋白质均无同源性。当将高度纯化的朊病毒制剂聚集成杆状且含有约10(9.5) ID50 U/ml时直接测序，发现了相同的氨基酸序列。氨基酸序列的知识应有助于确定朊病毒的遗传起源和复制机制。
