PowerPoint ????????? - Protein Engineering

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http//altair.sci.hokudai.ac.jp/

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• ??????? NMR
• ??????? K.Wüthrich

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Protein Engineering ????????????
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JournalProtein Engineering
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?????(???)??? ????1???? ???? 46
(22X)(22Y) ???? ?10?(105) ???? 30? (3 x
109) ???1m ?????? ???(4 x 106)
???1.4mm ?????(???)??? ?? ??????60??(6 x
1013) ???????? (DNA?RNA??????)
????5???????????? ????(??)7x105 hr ???
????90??????????? ????(??) 0.3?0.7 hr
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DNA??????????????

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DNA???
???????(Base)???????????? A???? C???? G????
T???
3???????1?????????(???)
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DNA??????AT?G C??????????????
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DNA????????
1st 2nd 3rd
5'-end U C A G 3'end
U Phe Ser Tyr Cys U
U Phe Ser Tyr Cys C
U Leu Ser Stop Stop A
U Leu Ser Stop Trp G
C Leu Pro His Arg U
C Leu Pro His Arg C
C Leu Pro Gln Arg A
C Leu Pro Gln Arg G
A Ile Thr Asn Ser U
A Ile Thr Asn Ser C
A Ile Thr Lys Arg A
A Met Thr Lys Arg G
G Val Ala Asp Gly U
G Val Ala Asp Gly C
G Val Ala Glu Gly A
G Val Ala Glu Gly G
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?????NMR
?????????????????????(NMR)??????????(?????????????
)?????????????????????????????????
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??????????2002 NMR?MS
2002????????
MS
NMR
John B. Fenn
Koichi Tanaka
Kurt Wuthrich
1/4 of the prize
1/4 of the prize
1/2 of the prize
http//www.nobel.se/chemistry/laureates/2002/index
.html
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NMR????????
http//www.nobel.se/
1952
1952
1991
2002
Kurt Wuthrich (1938 - ) ETH, Switzerland Scripps
Research Inst., USA
Felix Bloch (1905 - 1983) Stanford University USA
Edward Mills Purcell (1912 - 1997) Harvard
University USA
Richard R. Ernst (1933 - ) ETH, Switzerland
???(1991) for his contributions to the
development of the methodology of high resolution
nuclear magnetic resonance (NMR) spectroscopy.
???(2002) for his development of NMR spectroscopy
for determining the three-dimensional structure
of biological macromolecules in solution
????(1952) for their development of new
methods for nuclear magnetic precision
measurements and discoveries in connection
therewith.
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?????Nuclear Magnetic Resonance NMR
1H, 13C, 15N, 31P
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NMR??????????
??????? ???30300 µl ??0.33 mM
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???????? 1 Tesla10000 Gauss
???
???
4 x 10-1 T
850 T
3 x 10-1 T
60 T
10-2 T
Sun
33 T
10-4 T
2 (25) T
3 x 10-5 T
NMR
2 T
3 x 10-10 T
MRI
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NMR????? MHz??GHz?
????(Tesla) 0.8 ??? (1957) 1.4 2.1 2.3 4.7
??? 6.3 7.0 8.2 11.7 14.1 17.6 18.8 (1998) 21.1
(2001) 23.4 24.6 (1997) T.A. Cross et al., J.
Magn. Reson., 125, 212 (1997).
1H?????(MHz) 32 CW? 60 90 100
FT? 200 270 300 350 400 600 750 800 900
1000 1046
21.1 T
21.6 T (2002)
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(No Transcript)
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??????NMR???
???? 9.4??? 1H?????400MHz ????18.8???
1H?????800MHz
?????(???????????????) 1H0?10 ppm, 13C0?20 ppm,
31P0?600 ppm
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NMR??????????????
???????1H-1H???????(COSY) ????????????????????????
?
COSY
??????1H-1H????(lt5Å)???(NOESY) ???????????????????
????
NOESY
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NMR??????????
????????????????????????NMR???????????????????????
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????NMR???????????? ????
??????????????????(23-230)?NMR???? (K. Wüthrich
et al., )
Proc. Natl. Acad. Sci. USA 97(1), 145-150 (2000).
Proc. Natl. Acad. Sci. USA 97(15), 8334-8339
(2000).
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??????gt??? DNA?????????????? ????????????? ??????
???????? ???????????????? ??????? NMR ???????
K.Wüthrich