Division of Biofunctional Chemistry
The Chemical Society of Japan
Vol. 22, No.3 (2007. 12. 3)l   c M  
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ðî EF-Tu 3G[Uâ¬´ é«G[UâpÏ2-íEF-Tu 3tÐÞ

B-'3<ª°41î20 »3«G[Uâ3å.<1@3¢Ü¶V`IcL^bA2<æD?$íEF-Tu 4£1G[UGLatRNAën5íVal-tRNA ValíAla-tRNAAlaíPhe-tRNAPheì2{º3Ñ|.Åz#A/x?
¸?B-'î&3';íEF-Tu 4G[UGLa tRNA 3G[UâoãE8/F0ÖØ!-1/îîDB-'6!Aî&B 1? EF-Tu é«3G[UâoãEÆ#A/4ª4$.
Aî!! EF-Tu 4G[Uâ¬´ª/34×Ò.AîUhlenbeck ?4G[UGLa tRNA
24G[Uâás/ tRNA ásA/«3/2©·!íG[Uâás4´1A tRNA ás
{"3G[UGLa tRNA L`cNën5íVal-tRNA ValíAla-tRNA Val íPhe-tRNA ValìEm*EF-Tu /3ÅzE§Ú!' 5)î&!-í4 EF-Tu 41@G[UâEà@:#A/2¨,'î-2-k67:C½`1B=16F tRNA  EF-Tu "avt¾R

½Ala  Val )+ EF-Tu "¨c{Q,½OK)

½tRNAAla  tRNA Val )+ EF-Tu "º'½Ala-tRNAAla  Val-tRNA Val  EF-Tu "avtb¾%+½k¨c{67:C½1B=³²Y tRNA ²Y

rm>DG7/^½1B=16F tRNA  EF-Tu"avt/fI[,¾½EF-Tu 1B=³T´"{¸k1B=³

EF-Tu -,$0­ª+½,½

(w+,¾À¾ EF-Tu ¸k1B=³±a{
½¸k1B=³ EF-Tu a/®# 6)¾¸k1B=³/~ tRNA  EF-Tua¡/n½e¿)/z¾e¿q BODIPY-FL-AF ¸k
1B=³½k1B=³#,uj¸k1B=³

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1pyrAla-tRNA  EF-Tu  E215 T´!% (eÀÁ)¾½e¿_¸k1B=³½¥¯"pV¸sd·¾y½

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BODIPY FLC5 -AF
HN

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BODIPY FL aminophenylalanine(BODIPY FL -AF)

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C O OH
COOH
H 2N

COOH
H2N

H2N

H2N

O
O

O
HN
F
N

F
B

F
N
N

F
B

N

L-2-anthrylalanine(2antAla) L-2-acridonylalanine(acdAla )L-9-anthrylalanine L-1-pyrenylalanine L-2-pyrenylalanine(9antAla)

4P  7 C D    E 0; L8 ?# + )I M# +) # %- 

(1pyrAla): N(2pyrAla)

B 3O

4Q     # +) IB 3 *$ &(MR NJ@5       F3. B=>GM2N#+)IB3*$&(H1 <74M9NO
MS NJ@5       B3,'-O   /K!"O
MT N 6A.   B3,'-O-3-÷ó EF-Tu #6FCá¨±#u
EF-Tu #6FCá¨±#u"$V1óEF-Tu #6FCá¨±5ÓUhlenbeck /$òÅ EF-Tu #6FCáÃjE8>@#u±~í5z+òu±".0¨±
8)1tk ó)ò½Í/"$ò<JL¨±´! EF-Tu u±T5U~É<JL¨±´ EF-Tu "ÁQ1ó ê

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½Í$ 2003 )#äòOx}#¤Ú[Æ#NF@9LAJ6#ËÕ¿#·»5óF@9LAJ6$ò`$¶ÀÎ#,!1ÀÎ"\®±º®§×,
0ò-ÀÎÙ$©¼#ßS:;?G-ËÕ¿51óF@9LAJ6#ËÕ¿"$é"«ËÕn{- tRNA|qòÝf´!Ì51"$ïa´!êó!

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#6FCáà_<JLRv#,#"$Ãj<JL¨±

#.!_{

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EF-Tu "¨´!£s5¿Ä´"¹1u±~í5Ñó#ÃòEF-Tu #6FCáÃjE8>@"£s#u±5cøó x1¢¥´ EF-Tu 5<JL¨±´ EF-Tu "u1

 9)ó



!éy¦6FCá5Ô1 EF-Tu u± T#UÒ   

Ö5Y"òPm$x!éy¦6FCá",1.! EF-Tu 5U1

6FCá¨±5­1u$ò¨±5h"1.0¾g"
EF-Tu 5U2#/òx!éy¦6FCá5Ô1 EF-Tu $U21ó

pöú5Ó1ò1pyrAla #.!6FCáâ®r EF-Tu "Ãj!¬°$òx

Xã6FCáÃjE8>@#ä#¼Tæ421ó¨" E215 - D216 !
1."Óµ´1ó

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1òpöû#."V% 1pyrAla !

,6FCáÃjE8>@]"i)1$1óÃòp÷"¸Û0òâ®r EF-Tu Ãju±T- D216 u±TòE215A/D216A u±T$Ãj! 1pyrAla - 9antAla ",òE215A
1."!ó)ò2/#u±T$y¦#6FCá#ÃjÏ,W1ó)  
#  ' "  &  !   %        $(/"$òÐ³Ù'#ZòE215A u±Toè 1pyrAlaò2pyrAla  DL-2-anthraquinonylalanine ".& D216A u±T5¯1".0òÐ³Ù'#îdªZ"bðpøñóòEF-Tu 6FC6:K tRNA #ÃjÏ#î$!

ò#))Ð³Ùjdª"Ã&4ó EF-Tu $6FC6:K tRNA ÃjJD=MG'Þ&ò6FC6:K

tRNA 5å3^Ñ!!/!#òÃjç!#$ìµ!#1ó

E216A/D216A u±T"$ò1pyrAla-tRNA - 9antAla-tRNA #ÃjÏ

-4-

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

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,¥Ywt

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EF-Tu D©Y

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?:5B=D©Y'X[*-,´
COOH

COOH

COOH

H 2N

H 2N

H2 N
O
O

L-1-pyrenylalanine

DL-2-anthraquinonylalanine!      

L-2-pyrenylalanine   

 



µ´/+
 

EF-Tu  E215A $

D216A `0U,%³¨)Y)!³-*°b1<8ª¥"lQVg+a°b1<8ªC}´)

°b1<8ª1<8ªY;367«N­j

³Su



^

³
$D,´K' E ( D ªt1<8ª0 A `),¯®V(³`), EF-Tu RN§"q±0,s

,´$³a' EF-Tu "Zt

&M°b1<8ª'c],&³)1<8ª0¥lQ,&<8ªY;3670{,

Tx

W1

s.´LCk,&³Jr*` EF-Tu °b1<8ª9?2B4>@0_(³-*HO0¢#´¤¦ 

nmae fy~pi/-w+³Gh²0zo\IAF

dE)!¬P£v¤$´|
1. Hohsaka, T., Ashizuka, Y., Murakami, H., Sisido, M. Incorporation of nonnatural amino acids into
streptavidin through in vitro frame-shift suppression. J. Am. Chem. Soc. 118, 9778 (1996)
2. Bain, J. D., Switzer, C., Chamberlin, A. R. & Benner, S. A. Ribosome-mediated incorporation of anon-standard amino acid into a peptide through expansion of the genetic code. Nature 356, 537 (1992)
3. Hirao, I., Ohtsuki, T., Fujiwara, T., Mitsui, T., Yokogawa, T., Okuni, T., Nakayama, H., Takio, K., Yabuki,
T., Kigawa, T., Kodama, K., Yokogawa, T., Nishikawa, K. & Yokoyama, S. An unnatural base pair forincorporating amino acid analogs into proteins. Nat. Biotechnol. 20, 177 (2002)
4. Dedkova, L. M., Fahmi, N. E., Golovine, S. Y. & Hecht, S. M. Enhanced D-amino acid incorporation into
protein by modified ribosomes. J. Am. Chem. Soc. 125, 6616 (2003)
5. LaRiviere, F. J., Wolfson, A. D., Uhlenbeck, O. C. Uniform binding of aminoacyl-tRNAs to EF-Tu bythermodynamic compensation. Science 294, 165 (2001)
6. Nakata, H., Ohtsuki, T., Abe, R., Hohsaka, T. & Sisido, M. Binding efficiency of EF-Tu to tRNAs charged
with nonnatural fluorescent amino acids. Anal Biochem 348, 321 (2006)
7. Nissen, P., Kjeldgaard, M., Thirup, S., Polekhina, G., Reshetnikova, L., Clark, B. F. & Nyborg, J. Crystal
structure of the ternary complex of Phe-tRNAPhe, EF-Tu, and a GTP analog. Science 270, 1464 (1995)
8. Dale, T., Sanderson, L. E., Uhlenbeck, O. C. The affinity of EF-Tu for an aminoacyl-tRNA is modulated by
the esterified amino acid. Biochemistry 43, 6159 (2004)
9. Sato, A., Watanabe, Y., Suzuki, T., Komiyama, M., Watanabe, K. & Ohtsuki, T. Identification of theresidues involved in the unique serine specificity of C. elegans mitochondrial EF-Tu2. Biochemistry 45,
10920 (2006)
10. Ohtsuki, T., Sato, A., Watanabe, Y. & Watanabe, K. A unique serine-specific EF-Tu found in nematodemitochondria. Nat. Struct. Biol. 9, 669 (2002)
11. Doi, Y., Ohtsuki, T., Shimizu, Y., Ueda, T. & Sisido, M. EF-Tu mutants expand amino acid tolerance ofprotein biosynthesis system. J. Am. Chem. Soc. 129, 14458 (2007)-5-

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1. T. Hattori, M. Umetsu, T. Nakanishi, K. Tsumoto, S. Ohara, H. Abe, M. Naito, R. Asano, T. Adschiri, andI. Kumagai, “Grafting of material-binding function into antibodies -Functionalization by peptide grafting-”,
Biochemical and Biophysical Research Communications, in press.
2. T. Hattori, M. Umetsu, T. Nakanishi, S. Ohara, H. Abe, I. Kumagai, in preparation.

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1.

B. L. Vallee and R. J. P. Williams, Proc. Natl. Acad. Sci. U. S. A., 59, 498 (1968).

2.

a) S. T. Prigge, A. S. Kolhekar, B. A. Eipper, R. E. Mains and L. M. Amzel, Science, 278, 1300 (1997).
b) B. L. K. Kauppi, E. Carredano, R. E. Parales, D. T. Gibson, H. Eklund, S. Ramaswamy, Structure, 6,571 (1998). c) D. T. Logan, X. D. Su, A. Aberg, K. Regnstrom, J. Hajdu, H. Eklund, P. Nordlund,
Structure, 4, 1053 (1996).

3.

a) L. S. Childers, K. Folting, L. L. Merritt, Jr., and W. E. Strieb, Acta Cryst., B31, 924 (1975). b) E.
Boschmann, L. M. Weinstock, and M. Carmack, Inorg. Chem., 13, 1297 (1974).

4.

a) N. Kitajima, K. Fujisawa, C. Fujimoto, Y. Moro-oka, S. Hashimoto, T. Kitagawa, K. Toriumi, K.
Tatsumi, A. Nakamura, J. Am. Chem. Soc., 114, 1277 (1992). b) N. Kitajima and Y. Moro-oka, Chem.
Rev., 94, 737 (1994). c) B. Hazes, K. A. Magnus, C. Bonaventura, J. Bonaventura, Z. Dauter, K. H.
Kalk, W. G. Hol, Protein Sci., 2, 597 (1993).

5.

a) J. A. Halfen, S. Mahapatra, E. C. Wilkinson, S. Kaderli, V. G. Young Jr, L. Que, Jr., A. D.
Zuberbühler, and W. B. Tolman, Science, 271, 1397 (1996). b) B. MaHedman, K. O. Hodgson, T. D.
P. Stack, J. Am. Chem. Soc., 121, 5583 (1999). c) M. Kodera, K. Katayama, Y. Tachi, K. Kano, S.
Hirota, S. Fujinami, M. Suzuki, J. Am. Chem. Soc., 121, 11006 (1999). d) P. L. Holland, C. J. Cramer,
E. C. Wilkinson, S. Mahapatra, K. R. Rodgers, S. Itoh, M. Taki, S. Fukuzumi, L. Que, Jr., and W. B.
Tolman, J. Am. Chem. Soc., 122, 792 (2000).

e) L, M. Mirica, X. Ottenwaelder, T. D. P. Stack, Chem.

Rev., 104, 1013 (2004). f) E. A. Lewis; W. B. Tolman, Chem. Rev., 104, 1047 (2004).
6.

a) Y. Funahashi, K. Nakaya, S. Hirota, and O. Yamauchi, Chem. Lett., 1172 (2000).

b) Y. Funahashi

7.

A. Messerschmidt, R. Ladenstein, R. Huber, M. Bolognesi, L. Avigliano, R. Petruzzelli, A. Rossi, A.

and H. Masuda, Nippon Kessho Gakkaishi, 45, 328-334 (2003).
Finazzi-Agro, J. Mol. Biol., 224, 179 (1992).

-13-

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Karimata, N. Sugimoto, J. Am. Chem. Soc. 128, 7957 (2006). 7) D. Miyoshi, S. Matsumura, S. Nakano, N.
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