Abstract：
:Botulinum neurotoxins (BoNTs) and tetanus toxin (TeNT) are the most potent toxins for humans and elicit unique pathologies due to their ability to traffic within motor neurons. BoNTs act locally within motor neurons to elicit flaccid paralysis, while retrograde TeNT traffics to inhibitory neurons within the central nervous system (CNS) to elicit spastic paralysis. BoNT and TeNT are dichain proteins linked by an interchain disulfide bond comprised of an N-terminal catalytic light chain (LC) and a C-terminal heavy chain (HC) that encodes an LC translocation domain (HCT) and a receptor-binding domain (HCR). LC translocation is the least understood property of toxin action, but it involves low pH, proteolysis, and an intact interchain disulfide bridge. Recently, Pirazzini et al. (FEBS Lett 587:150-155, 2013, http://dx.doi.org/10.1016/j.febslet.2012.11.007) observed that inhibitors of thioredoxin reductase (TrxR) blocked TeNT and BoNT action in cerebellar granular neurons. In the current study, an atoxic TeNT LC translocation reporter was engineered by fusing β-lactamase to the N terminus of TeNT [βlac-TeNT(RY)] to investigate LC translocation in primary cortical neurons and Neuro-2a cells. βlac-TeNT(RY) retained the interchain disulfide bond, showed ganglioside-dependent binding to neurons, required acidification to promote βlac translocation, and was sensitive to auranofin, an inhibitor of thioredoxin reductase. Mutation of βlac-TeNT(RY) at C439S and C467S eliminated the interchain disulfide bond and inhibited βlac translocation. These data support the requirement of an intact interchain disulfide for LC translocation and imply that disulfide reduction is a prerequisite for LC delivery into the host cytosol. The data also support a model that LC translocation proceeds from the C to the N terminus. βlac-TeNT(RY) is the first reporter system to measure translocation by an AB single-chain toxin in intact cells.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Zuverink M,Chen C,Przedpelski A,Blum FC,Barbieri JTdoi
10.1128/IAI.00477-15subject
Has Abstractpub_date
2015-07-01 00:00:00pages
2714-24issue
7eissn
0019-9567issn
1098-5522pii
IAI.00477-15journal_volume
83pub_type
杂志文章abstract：:Live or heat-killed cells of Streptococcus mutans specifically adhere to dextran previously synthesized on glass surfaces by the action of extracellular dextransucrase. ...
journal_title：Infection and immunity
pub_type： 杂志文章
doi：10.1128/IAI.9.4.764-765.1974
更新日期：1974-04-01 00:00:00
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doi：10.1128/IAI.64.8.2955-2961.1996
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journal_title：Infection and immunity
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doi：10.1128/IAI.62.12.5581-5586.1994
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journal_title：Infection and immunity
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更新日期：1992-08-01 00:00:00
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更新日期：1998-02-01 00:00:00
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doi：10.1128/IAI.32.2.609-613.1981
更新日期：1981-05-01 00:00:00
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doi：
更新日期：1999-04-01 00:00:00
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journal_title：Infection and immunity
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doi：10.1128/IAI.63.5.1975-1979.1995
更新日期：1995-05-01 00:00:00
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journal_title：Infection and immunity
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更新日期：1977-11-01 00:00:00
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journal_title：Infection and immunity
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更新日期：1994-04-01 00:00:00
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journal_title：Infection and immunity
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更新日期：1976-06-01 00:00:00
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journal_title：Infection and immunity
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