??????????? PADI4 ? ?????

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??????????? PADI4 ????????????????????????

• ??18?3?23?
• ??????????
• ?????????????
• ???????????????????????
• ????????????????? ????????????????
• ?? ?

2
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1. ?????PADI?????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ?????

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(No Transcript)
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7
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8
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9
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10
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11
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12
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??

1. ?????PADI????????????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ???????????????

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NOS?NO??
??????
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?????????? 20? ?????? ??9? (??????? 3
?(Arg,His,Tyr)) ???10?(???????)
????? ??tRNA?? ???????????
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????????????????????
??????
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?????
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??????????
PADI??
???????????
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• ????
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• ???
• ?????
• ???????
• ??????
• ??????

??????? ??????
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?????????????????????????
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??

1. ?????PADI????????????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ???????????????

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?????????
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Korean
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PADI4 gene structure and association-positive SNPs
EXONs
SNPs
Caucasians
Japanese
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K562??????mRNA?????
Vector pcDNA-DEST40
CMV promoter
PADI4 (susceptible and non susceptible type cDNA)
5UTR 25bp
3 UTR 218bp
???? K562???Transfection??48?????? 5ug/ml
actinomycin D?? ??RNA????TaqMan assay???PADI4
RNA???????18S rRNA (???), Neomycin???(transfection
??)???????????
S susceptible type N non-susceptible type
100

Ratio of gradiation ()
50
plt0.05 (t-test)
S
N
S
N
S
N
0h
2h
4h
25
PADI4????RA?????????????????? ??????????
Standard curve
????DNA, cDNA PADI4????padi4_89
(A/G)?????????????4???????????????genomic
DNA?RNA Probe Padi4_89 (A/G) genotyping?TaqMan
probe
Ratio of G A alleles
Ave. genomic DNA 0.99 Ave. cDNA (4samples) 1.26
RA???????????????????????????????1???????cDNA????1
.3???????
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?????? ???

• PADI4???RA??????
• ????gt??????
• ????????
• ??????SNPs / ?????SNPs

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??

1. ?????PADI????????????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ???????????????

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PADI????

• ???????????
• ?????????????????????????
• ??????????(Ca2??)
• ?????
• ?????
• ?????????

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PADI???????????????????
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????? PADI???????????????
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Ca 210-3M
??????? Ca 2???PADI???? ??????????
Ca 210-7M
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??????? monocytes and macrophages
Monocyte
Macrophage
Transcriptional regulation
Enzyme
RNA
DNA
RNA
DNA
Enzyme
PADI4
Enzyme-activational regulation
Translational regulation
PADI2
ER Vossenaar WJ vanVenrooij LAB Joosten et al.
Arthritis Rheum 2004
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?????????
?????Arg
???
s-????Arg
a-????Arg
???
???
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???
???
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?PADI??????

• ?????
• ???????????????????????????????

????????????? ???ONOFF??
???? ????????????? ????? ?????????????? ???? PRMT?
????????
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??????????????????
Arg ? ???Arg ?????????? ????Arg ???????????
???
???
???
????????ON-OFF?????? ?????????????????????
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????????????????(??)
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??????????????????
Histone Sm (Small nuclear ribonucleoproteins)
Autoantigen
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?????

• Fibrinogen????
• ATIII?????
• Myelin Basic Protein(MBP)???

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MBP??????????????????????????(1970????2005???)
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??????????PADI????????????????
???????????????????????????
????? ???????????
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???????????????PADI???????
RA3???RA???B?????????scFv(???????)????????????????
?????????????scFv????????????????????RA???????????
??????????????????????????????????????????????????
??????????????????????????????????????????????????
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(No Transcript)
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PADI4????
46
??

1. ?????PADI????????????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ???????????????

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?CCP???????
RA?????????????
?CCP??????
?????????
?CCP???????? 9? ?????
?RA(OAReactive arthritis)?? ???????????
?CCP????B???????
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HLATCR??????????????
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(No Transcript)
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Augmentation of T-cell response by citrullinated
peptides
J.Hill E Cairns et al. J Immunol 2003
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Shared epitope
?????????? ???????????
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HLA-SE
aCCP()
?
Multi-hit model
CCP()-RA
CCP(-)-RA
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?????????????
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3. What are Substrates of PADI4 and Epitopes of
anti-CCP Ab?
Immunoscreening of citrullinated epitopes of RA
autoantibodies
Yamanaka OM, Suzuki A, Yamamoto K et al. 2005
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(No Transcript)
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(No Transcript)
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??????????
??????????

• citd Fib
• citd Fillagrin
• anti-Sa
• anti-perinuclear
• cit'ed CI
• cit'ed CII
• Eukaryotic elongation Factor
• citd alpha Enolase

• cit'd Fib
• cit'd Fillagrin
• anti-Sa ?
• anti-perinuclear ?
• cit'ed CI ?
• cit'ed CII ?
• Eukaryotic elongation Factor ?
• citd alpha Enolase ?
• Hisrone ?
• Sm ?
• MBP ?

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??

1. ?????PADI????????????????????????????
2. ???
3. ??????(????)
4. PADI???????????
5. ????
6. ??????
7. ????
8. ?????????????
9. PADI?????????????
10. ??????
11. ??????
12. ???????????????

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Streptococcal cell wall-induced? ????????
CIA????????? ?????? PADI24??????
??????Fibrin ???
?????????????? ????????
???????????????? CIA???
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• SNP Research Center, RIKEN, Yokohama, Japan
• Lab. for Rheumatic DIseases
• Kazuhiko Yamamoto
• Akari Suzuki
• Yuta Kochi
• Mikako Mori
• Kyoko Kobayashi
• Miyako Yamanaka
• Emi Kanno
• Keiko Myouzen
• Akihiro Sekine
• Tatsuhiko Tsunoda
• Yusuke Nakamura
• Clinical Institutes of Collaboration
• University of Tokyo Hospitals

• Center for Genomic Medicine, Kyoto University,
  Kyoto, Japan
• Fumihiko Matsuda
• Shohei Chida
• Alexandre Vasilescu
• Hitomi Hiratani
• Sachiko Toyoda
• Justine Yovo-Vasilescu
• Chanavee Ratanajaraya
• Miki Kokubo
• Kenei Ohigashi
• Victor Renault
• Masao Yamaguchi
• Katsura Hirosawa