Abstract:
:A procedure was developed to purify dermonecrotic toxin (DNT) from a sonic extract of a serotype D strain of Pasteurella multocida. Sonic extract containing DNT was applied to a DEAE-Sephacel column and eluted by a linear gradient of NaCl. Upon rechromatographing, fractions with dermonecrotic activity for guinea pigs were applied on a second Sephacel column, and a pooled fraction with the toxic activity was filtered through a Sephadex G-200 column. Pooled fractions with the toxic activity were subjected to polyacrylamide disc gel electrophoresis (PAGE), and the toxic substance was eluted from each sliced gel. Eluted fractions with the toxic activity were rechromatographed on a second Sephadex G-200 column, and a pooled fraction with high dermonecrotic activity was referred to as a purified DNT. The activity of purified DNT was increased by 1,000 times, and the average yield was about 1.8%. The purified DNT was homogeneous as determined by Ouchterlony double immunodiffusion, crossed immunoelectrophoresis, and thin-layer isoelectric focusing in polyacrylamide gels and gave a single band on PAGE and sodium dodecyl sulfate-PAGE. The molecular weight of the toxin was ca. 160,000 as determined by sodium dodecyl sulfate-PAGE. The isoelectric point of the toxin was ca. 4.7 to 4.8. Amino acid analysis of the purified DNT revealed that the toxin was composed of characteristically high proportions of glutamic acid, aspartic acid, glycine, proline, alanine, and leucine. The minimal necrotizing dose of the toxin was about 1 ng of protein, and the 50% lethal dose per mouse was 0.2 micrograms. The purified DNT was heat labile and sensitive to inactivation by trypsin, Formalin, and glutaraldehyde.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Nakai T,Sawata A,Tsuji M,Samejima Y,Kume Kdoi
10.1128/IAI.46.2.429-434.1984subject
Has Abstractpub_date
1984-11-01 00:00:00pages
429-34issue
2eissn
0019-9567issn
1098-5522journal_volume
46pub_type
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journal_title:Infection and immunity
pub_type: 杂志文章
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journal_title:Infection and immunity
pub_type: 杂志文章
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doi:10.1128/IAI.00098-07
更新日期:2007-06-01 00:00:00
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journal_title:Infection and immunity
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.43.1.359-367.1984
更新日期:1984-01-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.32.3.1014-1023.1981
更新日期:1981-06-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.52.1.1-5.1986
更新日期:1986-04-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.55.12.3174-3180.1987
更新日期:1987-12-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.62.2.673-679.1994
更新日期:1994-02-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.23.2.253-259.1979
更新日期:1979-02-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/IAI.60.5.1813-1819.1992
更新日期:1992-05-01 00:00:00
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journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.62.8.3206-3212.1994
更新日期:1994-08-01 00:00:00
abstract::Pertussis toxin is secreted from Bordetella pertussis with the assistance of the Ptl transport system, a member of the type IV family of macromolecular transporters. The S1 subunit and the B oligomer combine to form the holotoxin prior to export from the bacterial cell, although the site of assembly is not known. To b...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/iai.70.3.1193-1201.2002
更新日期:2002-03-01 00:00:00
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pub_type: 杂志文章,评审
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更新日期:2018-05-22 00:00:00
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journal_title:Infection and immunity
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doi:10.1128/IAI.26.1.15-18.1979
更新日期:1979-10-01 00:00:00
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更新日期:1993-05-01 00:00:00
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doi:10.1128/IAI.44.1.124-129.1984
更新日期:1984-04-01 00:00:00
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更新日期:2004-09-01 00:00:00
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journal_title:Infection and immunity
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更新日期:1996-10-01 00:00:00
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journal_title:Infection and immunity
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更新日期:1999-10-01 00:00:00
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更新日期:1999-09-01 00:00:00
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更新日期:2007-09-01 00:00:00
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journal_title:Infection and immunity
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更新日期:2002-02-01 00:00:00
abstract::Female guinea pigs were immunized with viable or UV light-inactivated chlamydiae (agent of guinea pig inclusion conjunctivitis), belonging to the species Chlamydia psittaci, by intravenous, subcutaneous, oral, or ocular routes. All animals were then inoculated vaginally with viable chlamydiae to determine the extent o...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.58.8.2599-2605.1990
更新日期:1990-08-01 00:00:00
abstract::Strains of Cryptococcus neoformans vary in resistance to phagocytosis in vitro. The binding of isolated capsular polysaccharide (CPS) to a capsule-free mutant of C. neoformans confers resistance to phagocytosis. The importance of capsule composition to differences among strains in susceptibility to phagocytosis was ev...
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更新日期:1989-12-01 00:00:00
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更新日期:1995-11-01 00:00:00