189
Vol. 32, pp. 189195, 2004
Biacore 1
1
1
2
A B
AB =*u-.nm*F
%vF
!%8G*F:T-2wd456x2=BC7+
!"#
JKJ 456%xU=:Mry/7
$%&'()*+,!-. /01-
%QRViB(0]73z()f{
.)2 () 3$45678#
e=01]73|}72Em:>2
09 %&'%8:;<=>
Biacore nmῌ8G:Mry/7%
!
?@ A/7 $%B&'3
QR~$QR*F: >i%Em QR
CD# *F ka =8GQR*F kd3W7)*
E3F#GH!"
:#IJK(0: $%LMNO& Surface
J
Plasmon Resonance: SPR =PQ.RS'3
e=01Z[3
(J0)*+,:T--. U$V%&
k F k \ $S#!=:7
'3
+
EJ76WXCD#!=:;
.\%F3X#!=()Y\.
%
LS
]7 Biacore
.=(0
$V&'= /S'3$45678
#098#Y+01127Z
^_F``, 320:;<V%&'
J
%)*!"q2) ab\%p()+0cdT
) $V%&'= 3!%$[\K%
~$fge3?)f#
456:#I#!=3]7# ^_^3
`9=J) a6N=
Biacore %89 bc=:;
<=%&'d e=fghῌ
! 1 Biacore SPR )*"%g3J
$d%>i%?j!%$V%4563
0 ! 1 % A, B, C %h Biacore %)*i3
(J)2k%k>i /0456%@A3BC
*h#3]J0
#!=l:
%gh /0%
L j<%klmj3
%?j%D7ZEmk>2
$V%456 nm*F K AZ2
8G*F KD =J)oH-.
!
kJ)2=+Y\.
..%)*n
%
L3J)2 $%S
$V%&':<I'%"JK()Lp
%<Inh mjL
%o
J KqMrs7Z=2(0NO:P17Z
pnh="qJ)2 /0
%op%nh
/0 A = B 456:Z = ABAB
rstuS#¡¢v()w
%QR: ABAB %QR3St()2
R%xy£RSPR £R¤¥3¦§# !%
%Em8G*F KD KD
SPR £R%nh3z{4¨©
6=*
!=3]7#
1 |ª}
2 ~«U¬
H ®
¯°W ±² ³
U¬ ´]µ,h $$8 ¶µ· ¸µ¹º»
T¼ ^_ F`
57
ª«
190
1
¬
®¯°
Biacore " SPR ῌSurface Plasmon Resonance῍ + A, B, C ¡¢£$%&'( Biacore ';bc῍ ¤¥
V$῍ ¦D`§Hq¨q©¦`aZdLῌ
῍ Biacore
)yrsdaῌ
ῌ
1 C $89:;>?[
!"#ῌ Biacore $῍ %
\'V]%^_`abcf῍ &'()*+,-./῎+01῏῍ -.
{|bc-./89:;
/23456789:;῎#<=+
Aarsdaῌ
01῏>?@A῍ %&'(BC)
Y"#῍ Biacore $ 2 xs2345
DEFG῍ H%&'(BC
v$
ZfBLYZῌ p῍
IJKFGLῌ HIJKM,NO
YYd"f῍ zhvk
%&'(BCPQR;S;-TR)D
ῌ
EM,῍ UOVWX
Biacore LYZῌ
1 A $-./*+,%&'(
Biacore "xs2345$῍
[\'V]%^_`abcd
1῏ ῌ 2῏ ef῍ ghL$ijklm9
"#$%&'() 3῏
H
!
Rn9oM,$WXpqakaῌ 1 B
4῏ *#+,qῌ Y
$῍ A bc-./23456789
7a
:;>?rsdaῌ $
1῏ ῌ
Lῌ
tuv-./234589:;wE
Biacore 5L01$῍ %BC
x)yd῍ zh{|bc}Y~
*+L01῎-./῏῍ 2345+
58
Biacore .X3$oOP
k6
191
῎
῏
῍ I$oOP
ῌ 1ῑ3῏ 55fy(,
SAῌ
!῍ "#
$
T2A55fy(,;῍ Biacore .
%&'()*+,-./01
KUt HBS ¡ ῌ10 mM HEPES pH
2ῌ
7.4, 150 mM NaCl, 3 mM EDTA, 0.005ῐ Surfac-
1ῑ1῏ 3456
tant P 20῍ ¢
mC῍ !£¤
758,9:3456;῍ <
_¥ ¦῍
῍ §V>5M.N¨;¦
=>?6'@AB$:CDE
;G©ῌ mC῍ 0.22 mm ()
@' DNA῍ RNA ABF$G῍ *+,-M.NWª;J2ῌ
HI
1ῑ4῏ «XC¡
῍ JDAKL$9
:M.NOP
/_0,1AB
!;QE῍ R
VW&'
"#'SABT$%U
!;῍ OP
¬KY!RY2ῌ m
!X2ῌ ();2῍ 3456
C;῍ OP
A;Y*AZK+[\2K῍
®῍ 758,?yzdC
90ῐ ]V^,-Gῌ .῍ /_0,"
C758,?yz«X2
!
12!"#;῍ 3456`a
ῌ «XC¡;῍
8,' pH AB£¤
Ab%$'3456$3c4ABd
G:t¯
Cef*Agh5i῍ !djk6
3456G;ῌ°t¯῍ DdC
lmC27n2ῌ
± [C t¯£¤ \]AAE
1ῑ2῏ Aῌ
ABZ22῍ 9:
2῏ 8,;$o9:;pq῍ !
<2./rAῌ =s;>?tk1
758,?yz`34569:;
KD @
u8,DE!v!-Gῌ
Eῌ 758,^²_dC
Biacore AE=w5xef*;῍ 75
`a9:bc῍ 2ED¥!3456P
8,?yz{BCD|EFG:
.r2d³$^9:
ῌ ./"};῍ 758,?yz9
3456~d³$efoa29
3456CD
dC.ef
:bc2ῌ
*G:῍ CD~|EF
2ῑ1῏ 758,?yz
Ee
®῍ <2
f*G:H:WAῌ u
34569:758,?yz; CM5,
ef*;*j῍ CDL῍ SA, NTA ABL2K῍ ~´.\
@8,῍
8,῍ G
A.gµ/ῌ
ῌ pH AB.KG:
῎1῏ 758,?yz CM5
/῍ $oIdC!
¶*·e¸?*¹x
5
3x
;῍ *jJ2ῌ
*jghK0
!
µ®῍ 5¶yz35f./345
I$oOP
!;῍ .2
69ῌ Q῎A34569
ῌ K῍ L6'*-AB.K
!῍ º Biacore M¢
CD Biacore 55fy(,M
8,?yz2ῌ
ῌ K῍ 758,?yz`
῎2῏ 758,?yz SA
N.r2Ogh
!P"
t75
758,?yz CM5 x
=z
»
#[\2ῌ =sP"#;῍ 3456
¼55¶yz35fdCt¯
9:7*MQ῍ +,*
ῌ »>?5:3456x
=z
»¼5-
5ABy5f
»>?5dC758,?yz9:
34569:
mCῌ AE
(,7*'R;:
(,7*[\
¢
758f./
ῌ
ῌ3῍ 758,?yz NTA
59
2
º»
192
^
_`¼a
CM5 NTA nitriloto-
#}x~ῌ
ri-acetic acid: II. 4,-()5@
ῌ !
Q_῍ `a8,-C6+
%&'
78k῍ #
<6GI4
"#$ Ni
2
()&'*+ῌ
+}x~ῌ
22 III. wOw+klT῍ bkO
,-
./0&'῍ /HE1w+()+[Ix
CM5 12
3415
6
~῍ 9:ῌ
789:;()+
IV. ;\<x=',-I&'*
9῍ <=>?@ABῌ
:ῌ
12
310CD)
3 ῏῍῎ῒῌῐῑ
N[-3-41/EF112G
Biacore >o4T῍ <=?
--EHE- NHS I N--E
3JKH
@ABῌ PB῍ ,-(
LG- EDC MN῍ )@PT6G\kl
12
310CD)&'5
6O
A)+78xῌ
H NHS 0+ῌ
31 >oB
()B,-86G 2 .
,-
CD)7P',-N
xb+lQ῍ CDx8
&'῍ ,-()@Qῌ ,
lBPklD4+
-()R'6STUV+ῌ
ῌ
WCD0XE
32 7D<E
1M Y
/1-ZL!+[I
2 .u>o+ῌF
8R'῍ 7CD0XE
+ῌ
_7D<E@Qῌ [B&'T
\
G 4H¡#'^¢+ῌ
XE
7]^῍ \
33 I<4
_`aN!+ῌ `aN!89῍
?J$b+,-
&
()&῍ 6G&'῍ @
,-῍ (),-defOgh&'
?J$I<£¤+ῌ [TI<
Pijk,-klmn+ῌ
¥K?J$P'¦_§@¨oh
5
6OHbc
&῍ [§@¨?J$I<£¤+ῌ
23 e1
34 KL©J$Hª
,-
.!e10%&'()+
Biacore 3«ST;M¬78I&kP
ῌ "oTIp
q#$ῌ [[T῍ rsTt%+ῌ
_῍ J$xNOkPQ®+
24 u:&'
RSῌ [Ta&'῍ ¯°
u:&'I&'T῍ 1 ()
KJ$L©J$Hª7+
vve),-
[I:ῌ
SA῍ 2 ()&w()* IgG w
[8QkHªT῍ 1 "ox±U
+()*,-w+&'+῍ 3 ()&
Vu4~[I῍ 2 W²X"ox~
w GST w+ GST .$&'+῍
[I῍ 3 5
4 NTA / Ni
2
%&'
`³Dx´P[I῍ 4 µYD
x¶kP[I῍ 5 0§+·Z~[I῍
!$&'+klxῌ
6 z[\D[Ikl
]
u:&'TyBz{x0P
&'P[Ix¸8x῍ Biacore T[B
His
9῍ <=$k1xῌ
]&'9῍ kHªz¹
I. ,-OH2+|D3
ῌ
60
Biacore uDX:('
4῏ F0'&?*&FNῌ /;῍
῍
@*;\῍ A*v<*0.!/0'
193
*0&o*Mῌ
!"#ῌ $%
Biacore &῍ Biacore '() ῌBIAevaluation῍ *
+,-.!/*0ῌ 1234
..*B/,;0CD>EF>Z
+5ῌ
e4:( Biacore u'G 1῏῍ 2῏
4ῒ1῏ 6789:8
ῌ
BIAevaluation *',;
)3HCD> mRNA & 5[ I
<=8>0῍ ?@A= 0 J7K!"LFK/
BCDE῍ 'FGHῌ I=
῍ DM\ N!v¡tOP¢DBQ
J῍ ?@A=KL)῍ MN&O8PQ
῎EF῏RS£FT¤Nῌ .
0NB&῍ R$S
7K/!UBEF>Z& eIF4
T0U:@VWῌ XY=)
E ῌeeukaryotic initiation factor 4E῍ !"῍ V
>Z![
EF>Z¥L¦(@!!vEF>ZWB
!E&
"\/0N
]῍ ' ^ G _ ` 0 * M ῌ 1 a XQEFRSNῌ n῍ 4EBP
ῌeIF4E binding protein῍ &EFe4Y§*
&XY=)!LbK/ 1 :
1 cd*B῍ Fef Langmuir model῍ M
M.!/ZNῌ ¨.*῍ EFe4Y
N& single-exponential binding model B!
§©0']῍ eIF4E ! 4EBP !
:(fN Biacore *
['
5g.!ῌ
,;ῌ
h&῍ analytical integration !
numerical integration ! # i ῌ i no῍ Xª«7L*
@¬7
global῍ local῍ separative fitting Fj/Mῌ $k
CM5 GST XBiDῌ i GST \
lmXY=)%&῍ no 1 : 1 B 4EBP ῌGST-4EBP῍ ῍ U]^
B9:8* global *LbK!®ῌ ¯XY=)!
4ῒ2῏ ,-ῌ
°_±* eIF4E ῍ ¨
'p!q(
*)/ Biacore
ῌ ²E῍ @¬7&
rs*tu-Fv!F;NFN
L6³´µ pH2.2 *`²Dῌ LbK*
ῌkdῑ1῍, ka )/w=YAF;NFNnox
M GST-4EBP ®,;E῍ °῎%
+ ῌ ) / , - * M B &῍
eIF4E ¶·aῌ b
. c2 )!yz7{)
¸cd 2 ¹ῌ R$S&῍ GST-
῎
global !$S)!z῏|/
4EBP eC· GST (N,;ῌ R$S
ῌ c2 )&01&I=J}?8!23 ῌῐ
T0U῎
῏ 3 ¹
2RU῍ !Fῌ ~S& 10RU 12F4N.
ῌ
*Mῌ n῍ yz7{)f0/5
3 a &® GST-4EBP °῎%
Fv*M.!x+/Mῌ
eIF4E B
῍ 3 b &
Ff0B&῍ )678&
7KXY{*M m7GTP !!v
Nῌ
eIF4E B῍ 3 c &R$S!
i local 9@K*
m7GTP eC· GTP fº eIF4E ,
N῍ %* kd )῍ ka )9
f0/
¹Nῌ
iN.!x+῍ 2 :(c
a, b, c ¨»
9
\
!῍ m7GTP fº b B& eIF4E ῎ON῏
Bῌ';ῌ
$%&῍ .<& BIAevaluation /
*
g¼h/·/ a c !
`i
=,;*῍ >0eu
N῍ .& m7GTP fºu; eIF4E 4EBP
61
·¸
194
ῐ2
¹º»¼
2
ῌBiacore @῍ 4EBP eIF4E rst
SPR k5
¦ GST ¦F§¨w
©{ª¨w«
DE῍ ¦FqC¬ 4EBP «
®¯5Iῌ R§¨w
©°
± eIF4E ῍ R !k5
ῌ
ῌ ῍ eIF4E ῌOFF῍ !"#
$%&'()$%῍ * 4
EBP +
,-.$%&%
/0῍ K D ῎k a ῏k d 1234+56 K D
$%&'784ῌ 9῍ *2
:; m7GTP <=ῐ 3 b GTP <=ῐ 3 c >?21%@A
῍ m7GTP eIF4E B
'C%DE eIF4E FGHIJK῍
eIF4E 4EBP LMN,OP
Qῌ
ῑ 1 ῍ ῐ 3 a, b, c R*S*
!@0
BIAevaluation @TU eIF4E 4EBP
VWPῌ ῑ 1 @0῍ ῐ 3 XY
C
Z[$ m7GTP <=
@ eIF4E 4
EBP %\῍ R*
]A+
ῐ3
56 KD I῍ ^90LMN6_C
`
῎%²³ 4EBP eIF4E pWq
rstI
aZ$ῌ *2Db
@0῍ 4EBP @
a ¯5I GST E 4EBP ´῎µ eIF4E
¶E !ῌ b m7GTP `
eIF4E ¶E῍ c GTP `
eIF4E ¶Eῌ
cd῍ eIF4E mRNA DEef`
g@hij*21%ῌ ^9
0῍ k5 Biacore k5
@0῍ lmn
o[%JFpWqrst()$῍ r
st
]lmnGHI9`u$
jv7ῌ
C719qC71῍ 9JF
7LM%1%.C
%
ῌ῏῎῍
Biacore %k57ῌ ,
wxyz{|}~
@῍ B5oC
2
Biacore k5῍ %.
lmn
rstlmn῎
`$῍ ¡¢£
ῌ5,*ῌ *2lmnrst
¤¥4@Aῌ
62
Biacore 4$56789:;<=>
῏1
GST-4EBP «#$¬῏® eIF4E ¯°±X²8
῍ ῍ &'ῌ ()
#$1%
῎῍ῐῌ
!"#$%
*+,-.3῏῍ 4῏῍ 5῏
/0
1῏ Miyoshi, H. et al.: J. Biochem. ῌTokyo῍, 126:
23&#$ῌ Biacore 4$56789
897ῐ904, 1999
:;<=>῍ ?@Aῌ BC
2῏ Youtani, T. et al.: IUBMB Life, 49: 27ῐ31,
DCEFGCHIEJKLMN@OP
QR
2000
S'TUVW$U῍ /XY<ZTT[
\T]^@_`CG
195
3῏ z{|}῍ ~῎῏῎1996῏GFf7
N$ῌ RI ab
4cdef9:;<=gh>
89:;<=(
῎ ῍
*c
4῏ ῍ ῎῏῎1998῏56f:;<=
$i῍ jklm4nahdef
GE>(
῎ FCῌ
:;<=obpq
HIC῍
rs$i
AtuOP%-nv$ῌ 56789:;<=
>OP%῍ /Xwxy$1%
5῏ ῍ ¡¢῎£¤῏῎1997῏GFf(
¥¦§C 1 O¨>῎ ©ª῍
23&
#$ῌ
63
© Copyright 2026 ExpyDoc