3.7 ᜪᐘણᠼᆢޑ០ނȐprecursorȑჹಒझࢥ܄ޑቹៜ
ϐΓॺ࣬ߞǴคፕࢂԋዕޑᜪᐘણᠼᆢ܈ࢂԋ୴᠄ᆫȐaggregateȑޑ
ೈқ፦ǴჹಒझԶقࣣࢂനڀࢥ܄ޑǴӢԜӧੰᏢ೯தୀෳࢂցԖ୴᠄ᆫ
ϐೈқ፦ޑ؈ᐘٰղۓࢂցᑡύኰઓسଏϯޑ੯ੰǶޔډന߈வӭਢٯ ύวǴځჴჹಒझനڀࢥ܄ޑᜪࠠϐࢂ pre-fibrillar aggregateȐΞᆀࣁᚇค ᠢޑᆫȐamorphous aggregateȑǵೈқ፦ϩηიȐprotein micelleȑ܈ࢂᠼᆢচࠠ
ȐprotofibrilȑȑԶߚԋዕޑᠼᆢǴձࢂଞჹ AEǵDsynuclein аϷ transthyretin ԶقǴΓൔύࣣࡰрځԐය܌ౢғޑᆫჹಒझࢂനڀࢥ܄ޑ134; 135; 136; 137;
138; 139
Ƕ
კ 3.9 ᆕᢀཥӝԋޑӭᴏ两ёૈޑၡ৩Ƕၡ৩ 1 ж߄ޑࢂ partially folded ᆶ native foldedޑރᄊǴӧݩΠೈқ፦ӛа native folded ԄӸӧǴฅԶऩࢂೈ
қ፦ౢғँᡂǵၸϯᏢঅႬ܈ࢂӸӧܭόᛙۓޑྋనΠ߾ೈқ፦а partially foldedԄӸӧǴѿౢғϼӭ partially folded ܈ denatured ޑೈқ፦ޑਔংǴ
ၸ proteasome ஒϐϩှǶऩԜమନᐒڋڙཞǴೈқ፦ԋόೕ߾ᆫ܈ࢂ
shifډၡ৩ 2 ޑၡጕǹၡ৩ 2 ܌وޑၡጕࢂӃԋ pre-fibrillar species നࡕӆᄌᄌ ߏԋԋዕޑᠼᆢȐၡ৩ 3ȑǶDANGER!߄ҢӧεϩޑਢٯύǴpre-fibrillar species
КԋዕޑᠼᆢڀԖၨଯޑࢥ܄ǴHeat shock proteinȐHspȑёаᔅշ෧Ͽ pre-fibrillar ޑԋǴٯӵஒ partially folded ޑೈқ፦҅ዴᄙ᠄ԋཥޑӭᴏ两܈ࢂஒό҅ዴᄙ
᠄ޑೈқ፦ϩှ6Ƕ
ԜѦҭԖЎ܌ၩ140Ǻஒӧᆫރೈқ፦Ԑය܌Ӄԋޑ pre-fibrillar species уΕਔჹಒझౢғࢥ܄Ǵ࣬ϸޑǴԋዕޑᠼᆢ߾ჹಒझ܌ԋޑࢥ܄্ၨ
λǴӢԜჹಒझ܌ౢғޑࢥ܄ࢂٰԾځ่ᄬౢғᆫ܌ᏤठǴԶߚӢࣁൂᡏᴏ两 Ȑmonomer polypeptideȑޑữ୷ለׇӈ܌ԋǴ೭ᆶԐӃךॺᇡࣁȨೈқ፦ޑ
ભ่ᄬёа،ۓΟભ่ᄬаϷғբҔȩޑᢀۺԖ܌рΕǶӢԜǴךॺགྷޕၰҁ ჴᡍύޑӚঁදҥܳᴏ两ჹಒझޑࢥ܄ቹៜࣁՖǴவॊޑჴᡍȐtime courseȑ
่݀ᡉҢǴӚᅿදҥܳᴏ两வൂᡏԋᜪᐘણᠼᆢ܌ሡޑਔ໔ନ catPrP คݤෳ
ளѦǴځᎩᴏ两ԋᜪᐘણᠼᆢ܌ሡਔ໔ऊ 40 λਔԿ 120 λਔόǴӢԜ
ᎦӧᎦనȐmediumȑ48 λਔࡕϐදҥܳᴏ两уΕ N2a ಒझਔǴჹಒझౢғޑ ࢥ܄ӵΠკȐკ 3.13ȑǴᒗޑಒझࢥ܄നமǴԛϐǴԶঊႵჹಒझޑ্നλǴ ӆٰࢂчऍεفജԛϐǴځᎩނᅿჹಒझޑ্࣬ৡόӭǹჹಒझࢥ܄ቹៜΟ ӜϩձࣁǺᒗǵаϷᒲႵǴฅځғߏᒨᅉයϩձࣁǺคݤෳளȐᒗȑǵ117.8±1.5 λਔȐȑаϷ 42.3±13.5 λਔȐᒲႵȑǶᒗᆶᒲႵදҥܳᴏ两ғߏᒨᅉයอǴӢ Ԝӧ࠻ྕΠᎦ 24 λਔࡕӆуΕಒझਔǴදҥܳᴏ两܌ԋޑ่ᄬ܈ࢂখӳ
ܰԋಒझࢥ܄ޑ pre-fibrillaǴฅԶჹܭғߏᒨᅉයၨߏޑᆶঊႵදҥܳᴏ 两ԶقǴݩ൩ၨᜤှǴᗨฅᆶঊႵදҥܳޑғߏᒨᅉය࣬ৡόӭȐϩձࣁ 117.8±1.5 λਔȐȑǵ118.9±4.9 λਔȐঊႵȑȑǴՠჹಒझӸࢲࠅԖ܌ৡ౦ Ȑ67.3±4.8 &Ȑȑǵ93.0±5.9 &ȐঊႵȑȑȐ߄ 3.5ȑǶ
bPrPcatPrP
dogPrPmdPrPmPrPhaPrPbvPrPlionPrP huPrP
pigPrPbuffer
0 20 40 60 80 100 120
Cell viability (%)
peptides
კ 3.10 ϯᏢӝԋޑදҥܳᴏ两ჹಒझࢥ܄ޑቹៜǶவಒझӸࢲளޕӚදҥܳ
ᴏ两ჹಒझޑࢥ܄࣬ৡόӭǴฅϝКၨр catPrP ڀၨமࢥ܄Զ haPrP ޑࢥ܄ന১Ƕ
߄ 3.9
Peptides lag time
Ȑhrȑ
Cell viability Ȑ%ȑ
bPrP 61.1±10 76.5±5.7
catPrP / bPrPV112M N.D 63.8±3.7
dogPrP / bPrPM129L 71.4±12*** 77.7±5.9 bvPrP / bPrPL138M 42.3±13.5** 69.7±6.7 mdPrP / bPrPS135N 72.2±5.5 85.9±6.1 pigPrP /bPrPN143S 117.8±1.5 67.3±4.8 lionPrP
(V112M,S135N) 51.2±14.3 73.4±6.0
mPrP
(M109L,L138M) 65.5±6.8*** 77.2±4.3
haPrP
(V112M,L138M,I139M) 118.9±4.9*** 93.0±5.9
huPrP
(V112M,L138I,N143S) 61.8±8.4*** 78.9±6.9
ഢຏΚp<0.05ʽΙp<0.01**Ιp<0.001***
߄ 3.9 Ӛঁදҥܳᴏ两ځғߏᒨᅉයᆶჹಒझࢥ܄ޑቹៜǶჹಒझࢥ܄ቹៜന εޑΟӜࣁᒗǵᆶᒲႵǴԶჹಒझࢥ܄ቹៜനλޑࣁঊႵǴځᎩނᅿჹಒझ ޑቹៜ࣬߈Ƕ
ಃѤകʳ ่݀ᆶፕ!
!
!!! TSE ္܌ᇥޑᅿࡀምࢂࡰǺނᅿஒ TSE ੯ੰགࢉќނᅿਔǴག
ࢉޑނᅿჹԜ੯ੰԖܢל܄ǶԶ܌ᒏޑܢל܄൩ࢂǺӧ TSE agent གࢉཥޑނᅿ ਔǴ၀ނᅿಃжևԖၨߏޑᎦਔ໔ǴԶ೭ঁਔ໔ࣗԿຬၸނᅿޑғڮຼ
යǶՠࢂ agent ᒿኧж౽ਔǴ၀ނᅿౢғᔈ܄٠ЪᕭอᎦਔ໔ޔၲ
ᛙۓރᄊǶԾவम୯ᆶऍ୯ᛈว BSE ᆶ CWDǴךॺ࣬ߞǴBSE ᆶΓᜪ vCJD ࢂ
࣬ᜢޑǴΨࢂவٗਔংω໒ۈख़ຎ TSE ჹځдނᅿགࢉޑᅿࡀምୢᚒǶӧނ ᅿགࢉ prion agent ਔǴdonor ᆶ receptor ޑ PrP ữ୷ለׇӈϐ࣬՟ࡋӧᙯඤၸำ ύתᄽख़ाفՅǶPrP ữ୷ለׇӈ࣬՟ࡋଯǴ߾ԋ PrP-res ޑਏӳǴΨ൩ࢂ
PrP-res ၨזಕᑈԶౢғᖏ੯ੰǴϸϐ PrP ữ୷ለׇӈ࣬՟ࡋեޑ၉ȐЇ
ਏ݀ৡȑǴᏤठ PrP-res όܰԋԶफ़եΑੰวၸำǶ
ӧҁԛჴᡍύךॺᄒڗჹܭᜪᐘણԋ࣬ख़ाޑࢤȐ࣬ܭΓᜪදҥܳ
ׇӈ 108-145*ࣁኳࠠǴନΑۓໆࢉࡀምѦǴ٠׳ΑှӧᅿӸӧΠߦ
PrPC่ᄬޑᙯᡂၸำύਸޑữ୷ለ܌ٰޑቹៜǺM112 ӢࣁуೲΑԋਡբҔ
٬ள׳ӭޑE-PrP ёٮЇਔ٬ҔǴӢԜߦ PrP ่ᄬޑᙯᡂǴS143ǵM139
ۯᒨԋਡբҔǴԶуೲᆶۯᒨٿӢηӸӧ࣬ӕ peptide ਔǴ࣬ϕܢᎍ ځਏ݀ǴӢԜঊႵȐhaPrPȑᆶΓᜪȐhuPrPȑޑғߏᒨᅉයᆶФȐbPrPȑ࣬߈Ƕ КၨФᆶജᅿǴϩձჹӚނᅿՉЇࡀምวځ่݀ᜪ՟ǴჹܭᒲႵǵλ ႵǵᒗаϷྰηނᅿǴவΓޑჴᡍ൩ςჴڙډ BSE ܈ࢂ CWD ޑག
ࢉ 18; 121ǴӢԜჹ೭٤ނᅿޑЇਏӳǴᅿࡀምҭ᪲ǴձࢂჹᒗٰᇥǴϐ
ԖࣴزᡉҢ BSE К CWD ׳ܰЇଆᒗགࢉ TSE ੯ੰǴԶҗЇբҔҭว
ӕኬޑ่݀ǹԶჹކǵٰᇥǴࢂҢࣁჹ BSE ܈ࢂ CWD ԖܢלΚޑނᅿǴ ӢԜЇਏ݀ৡǴᅿࡀምଯǶԜѦǴคፕࢂФჹജՉЇჴᡍ܈ࢂജᅿ ჹФՉЇǴࣣว۶Ԝ໔ޑᅿࡀምࡐଯǴӢԜճҔ oral БԄၠᅿགࢉޑ
ёૈ܄ࡐեǹКၨٯѦޑࢂ௶ճ٥ঊႵǴ௶ճ٥ঊႵᗨฅڙډ BSE གࢉǴՠ
CWD གࢉਔࠅӢӸӧόӕ strain ԶቹៜځགࢉਏȐऩ strain ӧ 135 Տࣁ SerǴ߾Їࡀምࣁ 1Ǻ2Ǵऩ strain ӧ 135 Տࣁ AsnǴ߾Їࡀምࣁ 1Ǻ8ȑǹჹ ܭΓᜪԶقǴςวӢӞΑԖئФੱޑФԺԶᏤठᑡ vCJDǴவЇჴᡍ
ჴФჹΓϐ໔ޑᅿࡀም٠όଯǴՠԿϞࠅۘ҂วӢӞΑڙ CWD གࢉޑജ ԺԶᑡΓᜪ TSE ੯ੰǴԶךॺޑჴᡍΨวജჹΓϐ໔ޑଯᅿࡀምǶа bPrP ࣁᅿՉЇਔǴM138 फ़եЇࡀምǴL129ǵN135ǵM139 аϷ S143 ࣣ
ቚуЇࡀምǴӢԜफ़եᆶቚуٿӢηӕਔӸӧ࣬ӕ peptide ਔǴځਏ݀
ύکǴႽࢂΓᜪȐhuPrPȑǴԜѦ bPrP ჹ dogPrP ᆶ pigPrP ޑЇࡀምϩձࣁ 4 ७ᆶ 8 ७Ǵ൩ϩηቫԛԶقǴBSE ၨᜤၠຫᅿࡀምࢉ๏ކᆶǶԶа mdPrPࣁᅿՉЇਔǴmdPrP ჹ haPrP ޑᅿࡀምࣁ 8 ७ǴӢԜ CWD ၨᜤ
ၠຫᅿࡀምࢉ๏ঊႵǴԜѦ mdPrP ჹ huPrP ޑᅿࡀምࣁ 16 ७ᡉҢ CWD К BSE ׳ᜤࢉ๏ΓᜪǶஒӝԋޑදҥܳᴏ两ځԾว܄ԋᜪᐘણΚᏢаϷ Їࡀም่݀ԋკ߄Ȑ߄ 4.1ȑǶ
ԶவΚᏢྋှࡋჴᡍύวǴᗨฅҁጇፕЎύ܌Ԗӝԋޑදҥܳᴏ两ྋှ
ࡋࣣࡐӳǴՠӚදҥܳᴏ两ϝӸӧᠼᆢྋనύ҂ԋᜪᐘણᠼᆢޑ PrPScൂᡏᐚ ࡋᆶځữ୷ለϯᏢឦ܄٠ό࣬಄Ъᆶ lag time کЇࡀምҭค࣬ᜢ܄ǶԜѦǴӚ දҥܳᴏ两ჹಒझࢥ܄ޑբҔ࣬ৡόӭǴ٠คܴᡉձǶ
ჹදҥܳೈқٰᇥǴữ୷ᆄޑׇӈ 1-94 ჹܭ PrP-res ޑԋόڀख़ा܄ǴӢ ࣁ೭ࢤୱ࣬ჹచදҥܳೈқٰᇥڀԖቸ܄ЪόڀԖҺՖ่ᄬ܄ǴฅԶࠅёૈ
ੋϷϩη໔Ȑinter-ȑ܈ϩηϣȐintra-ȑჹܭೈқ፦ύЈȐprotein coreȑǵಒझ߄ य़аϷϺฅଛᡏȐnatural ligandȑ໔ޑբҔǹฅԶׇӈ 106-147 ࠅࢂᄬᙯᡂ୴
᠄ԋᜪᐘણᠼᆢЬाวғޑୱǶӧεӭኧޑࣴزύ܌٬ҔޑࢂλТࢤޑදҥ
ܳᴏ两ԶჹܭදҥܳೈқޑځдୱࢂցԖϟΕᜪᐘણᠼᆢޑԋࠅόమཱǴவ ҁჴᡍ่݀Ψளޕൂữ୷ለޑॄय़ቹៜȐᜪᐘણԋਏᆶЇࡀምȑёа
҂ٰӵૈஒࣴزߏࡋۯ՜Ǵஒޑόஒ߳ज़ܭׇӈޑ࣬՟ࡋჹ PrP-res ޑ ቹៜǴԶࢂ൩ᡏදҥܳೈқޑΟભ่ᄬࣴزǶ!
߄ 4.1 දҥܳᴏ两ځԾว܄ԋᜪᐘણΚᏢǵΚᏢྋှࡋаϷЇࡀም
Peptides lag time (hr) using bPrP
fibrils as seeds fibrils as
seeds bPrP 61.1±10 1.95±0.33 1 4 mdPrP/bPrPS135N 72.2±5.5 1.62±0.24 8 1
One-residue different
catPrP / bPrPV112M N.D 1.65±0.11 1 2 dogPrP / bPrPM129L 71.4±12 1.19±0.07*** 4 4 bvPrP / bPrPL138M 42.3±13.5* 1.39±0.01** 0.5 1 pigPrP /bPrPN143S 117.8±1.5*** 2.79±0.23*** 8 4
Two-residue different lionPrP
(V112M,S135N) 51.2±14.3 1.63±0.06 2 2
mPrP
(M109L,L138M) 65.5±6.8 1.09±0.02*** 2 2
Three-residue different haPrP
(V112M,L138M,I139M) 118.9±4.9*** 2.95±0.59*** 2 8
huPrP
(V112M,L138I,N143S) 61.8±8.4 1.16±0.07*** 2 16 ഢຏΚp<0.05ʽΙp<0.01**Ιp<0.001***
N.DΚnot-detectable
bPrPcatPrdogPrPbvPrPmdPrPlionPrPmPrPpigPrPhuPrPhaPrPP 0
2 4 6 8 10 12 14 16
seeding barrier
target peptides
mdPrP as seed bPrP as seed
კ 4.1 а bPrP ܈ mdPrP ࣁᅿჹӚނᅿදҥܳᴏ两ϐЇࡀምǶคፕࢂа bPrP
܈ mPrP ࣁᅿਔǴჹ dogPrP ᆶ pigPrP ϐЇࡀምࣣଯȐ1:4ȑǴฅԶа mdPrP ࣁᅿਔǴჹ haPrP ᆶ huPrP ϐЇࡀምȐϩձࣁ 1:8 ᆶ 1:16ȑКа bPrP ࣁᅿ ਔଯȐϩձࣁ 1:2 ᆶ 1:2ȑǶ
ୖԵЎ
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