Nuclease treatment results in high specific purification of Creutzfeldt-Jakob disease infectivity with a density characteristic of nucleic acid-protein complexes.
Representative preparations of partially purified Creutzfeldt-Jakob disease (CJD), including disaggregated density gradient fractions, were treated with a variety of nucleases. RNases as well as exhaustive digestions with micrococcal nuclease did not significantly diminish infectivity, but resulted in an approximately 7,000-fold specific purification of infectivity with respect to nucleic acid. Protected nucleic acids included species of up to 2,000 bases in length. After nuclease treatment, infectivity co-migrated with nucleic acid-protein complexes at a density of 1.27 g/cm3 in sucrose. Substantial specific protein purification were also achieved in the gradient step (approximately 11,000-fold), where 70% the host Gp34 ("prion protein") as well as other free proteins separated from infectivity. These CJD purifications are better than those previously attained in scrapie, and may be useful for further studies of non-host protein and nucleic acid species. The data are consistent with the hypothesis that CJD-like agents are composed of nucleic acid-protein complexes.
Sklaviadis
T
T
Yale University Medical School, New Haven, Connecticut.
Akowitz
A
A
Manuelidis
E E
EE
Manuelidis
L
L
D000818:Animals; D002500:Centrifugation, Isopycnic; D007562:Creutzfeldt-Jakob Syndrome / Q000382:microbiology; D006224:Cricetinae; D003851:Deoxyribonucleases / Q000378:metabolism*; D009696:Nucleic Acids / Q000302:isolation & purification / Q000378:metabolism; D018620:PrPSc Proteins; D011328:Prions / Q000302:isolation & purification* / Q000378:metabolism / Q000472:pathogenicity; D012260:Ribonucleases / Q000378:metabolism*; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism
D009696:Nucleic Acids; D018620:PrPSc Proteins; D011328:Prions; D014764:Viral Proteins; D003851:Deoxyribonucleases; D012260:Ribonucleases