Abstract：
:Purification of the scrapie agent by methods using digestion with proteinase K yields a protein product, PrP-27-30, with an apparent mass of 27-30 kDa (D. C. Bolton et al. (1982) Science 218, 1309-1311; S. B. Prusiner et al. (1982) Biochemistry 21, 6942-6950). In contrast, a 33-37 kDa glycoprotein, HaSp33-37, was the major protein component isolated from scrapie-affected hamster brain by a procedure that did not use protease digestion. The purified fractions containing HaSp33-37 had greater than 10(11) LD50 units of the scrapie agent per milligram of protein. Proteinase K digestion of HaSp33-37 gave a product indistinguishable from PrP-27-30 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The amino acid sequence of the first 22 residues of HaSp33-37 was determined. The sequence coincided with that predicted for the N-terminus of the precursor to PrP-27-30 (K. Basler et al. (1986) Cell 46, 417-428; N. K. Robakis et al. (1986) Proc. Natl. Acad. Sci. USA 83, 6377-6381) after processing by signal protease. HaSp33-37 was digested with N alpha-tosyl-L-phenylalanine chloromethyl ketone-trypsin to produce a 29-32 kDa protein fragment; following digestion this fraction retained complete biological activity. The amino terminal sequence of the 29-32 kDa protein corresponded to a position intermediate between the amino termini of HaSp33-37 and PrP-27-30. We conclude that HaSp33-37 is the intact form of the scrapie agent protein and that PrP-27-30 is produced by proteinase K degradation when this enzyme is introduced during isolation of the scrapie agent.
journal_name
Arch Biochem Biophysjournal_title
Archives of biochemistry and biophysicsauthors
Bolton DC,Bendheim PE,Marmorstein AD,Potempska Adoi
10.1016/0003-9861(87)90380-8subject
Has Abstractpub_date
1987-11-01 00:00:00pages
579-90issue
2eissn
0003-9861issn
1096-0384pii
0003-9861(87)90380-8journal_volume
258pub_type
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更新日期：1999-09-01 00:00:00
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pub_type： 杂志文章
doi：10.1006/abbi.1994.1407
更新日期：1994-10-01 00:00:00
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pub_type： 杂志文章
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pub_type： 杂志文章,评审
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pub_type： 杂志文章
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更新日期：1984-12-01 00:00:00
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pub_type： 杂志文章
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更新日期：1999-07-15 00:00:00
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journal_title：Archives of biochemistry and biophysics
pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：1994-02-15 00:00:00
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pub_type： 杂志文章
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更新日期：2007-02-15 00:00:00
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journal_title：Archives of biochemistry and biophysics
pub_type： 杂志文章
doi：10.1006/abbi.1993.1086
更新日期：1993-02-01 00:00:00
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更新日期：2003-01-01 00:00:00
