Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme
Leucyl-tRNA synthetase from Thermus thermophilus (LeuRSTT) was purified to homogeneity using a five-step purification procedure. The enzyme was characterized and crystallized. Molecular mass determinations of the native and denatured proteins indicate monomeric structure of LeuRSTT with the molecula...
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Інститут молекулярної біології і генетики НАН України
2001
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| Cite this: | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme / A.D. Yaremchuk, O.I. Gudzera, S.P. Egorova, D.I. Rozhko, I.A. Kriklivy, M.A. Tukalo // Біополімери і клітина. — 2001. — Т. 17, № 3. — С. 216-220. — Бібліогр.: 11 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1860139398328221696 |
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| author | Yaremchuk, A.D. Gudzera, O.I. Egorova, S.P. Rozhko, D.I. Kriklivy, I.A. Tukalo, M.A. |
| author_facet | Yaremchuk, A.D. Gudzera, O.I. Egorova, S.P. Rozhko, D.I. Kriklivy, I.A. Tukalo, M.A. |
| citation_txt | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme / A.D. Yaremchuk, O.I. Gudzera, S.P. Egorova, D.I. Rozhko, I.A. Kriklivy, M.A. Tukalo // Біополімери і клітина. — 2001. — Т. 17, № 3. — С. 216-220. — Бібліогр.: 11 назв. — англ. |
| collection | DSpace DC |
| container_title | Біополімери і клітина |
| description | Leucyl-tRNA synthetase from Thermus thermophilus (LeuRSTT) was purified to homogeneity using a five-step purification procedure. The enzyme was characterized and crystallized. Molecular mass determinations of the native and denatured proteins indicate monomeric structure of LeuRSTT with the molecular mass of about 101 kDa. The protein obtained is remarkably thermostable and retains 97 % of its initial aminoacylation activity after 1 hour of incubation at 88 °C. Crystals of LeuRSTT were obtained from ammonium sulfate solution by the vapour diffusion techniques. The crystals quality was improved by crystallization from the precipitate.
Лейцил-тРНК синтетаза из Thermus thermophilus (ЛейРСТТ) выделена в гомогенном состоянии с использованием пяти стадий очистки. Фермент охарактеризован и получены его кристаллы. Определена молекулярная масса нашивного и денатурированного белка. Установлено, что ЛейРСТТ представляет собой мономер с молекулярной массой 101 кДа. Полученный фермент обладает значительной термостабильностью и сохраняет 97 % аминоацилирующей активности после инкубации в течение 1 ч при температуре 88 °С. Кристаллы ЛейРСТТ получены методом диффузии паров с использованием в качестве осадителя раствора сульфата аммония
Лейцил-тРНК синтетазу із Thermus thermophilus (ЛейРСТТ) виділено в гомогенному стані з використанням п'яти стадій очищення. Фермент охарактеризовано та отримано його кристали. Визначено молекулярну масу нативного і денатурованого білка. Встановлено, що ЛейРСТТ являє собою мономер з молекулярною масою 101 кДа. Отриманому ферментові притаманна значна термостабільність і він зберігає 97 % аміноацилюючої активності після інкубації протягом 1 год при температурі 88 ° С Кристали ЛейРСТТ одержано методом дифузії парів із використанням як осаджувана розчину сульфату амонію.
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ISSN 0233-7657. Біополімери і клітина. 2001. Т. 17. № З
Leucyl-tRNA synthetase from Thermus thermophilus.
Purification and some properties of the crystallizing
enzyme
A. D. Yaremchuk, О. I. Gudzera, S. P. Egorova, D . I. Rozhko,
I. A. Kriklivy, M. A. Tukalo
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine
150 Academician Zabolotnoho vul., Kyiv, 03143 , Ukraine
Leucyl-tRNA synthetase from Thermus thermophilus (LeuRSTT) was purified to homogeneity using a
five-step purification procedure. The enzyme was characterized and crystallized. Molecular mass
determinations of the native and denatured proteins indicate monomeric structure of LeuRSTT with the
molecular mass of about 101 kDa, The protein obtained is remarkably thermostable and retains 97 % of
its initial aminoacylation activity after 1 hour of incubation at 88 °С. Crystals of LeuRSTT were obtained
from ammonium sulfate solution by the vapour diffusion techniques. The crystals quality was improved by
crystallization from the precipitate.
Introduction. Aminoacyl-tRNA synthetases (aaRSs)
catalyze the specific esterification of an amino acid to
the З'-end of its cognate tRNA. Through a two-step
reaction, aaRSs use first ATP to form the activated
intermediate, aminoacyl-adenylate, and then transfer
the amino acid to the З'-end of their cognate tRNA.
The reaction is highly specific, although in some
cases, to ensure the fidelity of the genetic code
translation, the aaRS relies on an editing activity to
hydrolyze misactivated amino acids which are similar
to the cognate amino acid [1, 2] . The family of
twenty aaRSs are divided into two distinct classes of
ten enzymes each on the basis of the primary and
tertiary structures information [3, 4 ]. Also, all tRNA
molecules can be classified into two types according
to the length of the variable arm. Type I tRNAs (most
of tRNA molecules) have a short variable arm com
prising four or five nucleotides. Type II tRNAs
(tRNA 1^ from eubacteria and organelles of lower
eukaryotes, tRNA U u and tRNA S e r) have a long va
riable arm composed of more than ten nucleotides.
Leucyl-tRNA synthetase (LeuRS) is a monomeric
class I enzyme which recognizes type II tRNA. It was
© A. D. YAREMCHUK, О. I. G U D Z E R A , S. P. EGOROVA,
D. I. ROZHKO, I. A. KRIKLIVY, M. A. TUKALO, 2001
shown by mutational studies that Escherichia coli
LeuRS recognized tRNA U u in a manner different from
the standard recognition mode [5] . Only the
discriminator base A73 is required for the base-
specific recognition by LeuRS while the anticodon, the
variable stem loop and the acceptor stem sequences
are not essential for the specific recognition by
LeuRS. To define the recognition mode and structural
identity elements of tRNA1*" by LeuRS as well as to
get detailed information on substrate specificity and
enzyme mechanism it is necessary to determine the
crystal structure of the enzyme alone and in the
complex with substrates. So far crystal structures of
sixteen from twenty different synthetases have now
been determined together with a variety of substrate
complexes [6, 8 ]. LeuRS is one of four aaRSs which
crystal structure is not known yet. Here, we report
the purification, some properties and crystallization of
LeuRSTT.
Materials and Methods. Materials, DEAE Sepha-
rose Fast Flow, Heparin Sepharose CL-6B were from
«Farmacia LKB Biotechnology» (Sweden). Hydro-
xylapatite was from «Віо-Rad» (USA). TSK-Gel HW-
65 from «Тоуо Soda MFG» (Japan). Polypepton and
yeast extract were from «Difco» (USA). Unfrac-
tionated tRNAs from E. colU protease inhibitor cock-
216
LEUCYL-tRNA S Y N T H E T A S E FROM T H E R M U S THERMOPHILUS
tail tablets (completeTM) and phenylmethylsulfonyl
fluoride were from «Boehringer Mannheim» (Ger
many). Protein markers for molecular mass deter
minations, HEPES and ATP were purchased from
«Sigma Chemical Со.» (USA). 1 4C-labelled leucine
was from «Amersham, UK» (Great Britain). Ammo
nium sulfate, polyethylene glycol, MES and bis-Tris-
propane were from «Fluka, Biochem.» (Switzerland).
Purification of T. thermophilus LeuRS and
tRNAUu. The T. thermophilus (strain HB8 or HB27)
cells were harvested by centrifugation, washed and
disrupted by a French Press in 100 mM Tris-HCl
buffer, containing 2 mM dithiothreitol, 0.1 mM
EDTA, 1 mM phenylmethylsulfonyl fluoride, com
pleteTM (1 tabl./25 ml extraction buffer), 5 mM
MgCl2 and 1 mM NaN 3 . The extract was centrifuged
for 2 hours at 105000 g and the supernatant was
subjected to 30—65 % ammonium sulfate fractio
nation. The precipitate containing LeuRS activity was
dissolved in 20 mM Tris-HCl buffer, pH 7.5 con
taining 5 mM MgCl2, 0.1 mM phenylmethylsulfonyl
fluoride, 2 mM DTT, 1 mM NaN 3 (buffer A),
dialyzed against buffer A and absorbed on a DEAE-
Sepharose column (5 x 55 cm) equilibrated with buff
er A. The LeuRS was eluted with 2 x 2.5 L linear
gradient of 0.03 M to 0.3 M sodium chloride in buffer
A. The fractions containing LeuRS activity were
salted out by ammonium sulfate (50 % saturation)
and applied to a Toyopearl HW-65 column (3 x
x 80 cm) equilibrated with 40 % of ammonium sul
fate in buffer A. The proteins were eluted with 2 x 2
L gradient of 40—10 % saturation ammonium sulfate
in buffer A. Fractions containing LeuRS activity were
pooled, dialyzed and chromatographed on a hyd-
roxylapatite column (3 x 40 cm) with 2 x 2 L potas
sium phosphate buffer linear gradient from 0.01 M to
0.25 M (pH 7.9). After dialysis in buffer A of the
active fractions the enzyme was loaded on a Heparin
Sepharose CL-6B column (1 x 40 cm). A 1.0 L linear
0—0.25 M KC1 gradient in buffer A was used to elute
the LeuRS. All steps of the enzyme purification were
carried out at 4 °С
T. thermophilus tRNA U u was purified from bulk
tRNA using BD-cellulose, anion-exchange DEAE
5PW (HPLC, «Весктап», USA) and C8 reverse-
phase chromatography (I. Krikliviy, D. Rozhko and
M. Tukalo, unpublished results).
Enzyme activity. The aminoacylation reaction mi
xture contained 50 mM Na-Hepes, pH 7.3, 15 mM
MgCl2, 30 mM KC1, 0.5 mg/ml bovine serum albumin
(BSA), 7 mM ATP, 0.1 mM DTT, 0.096 mM L-
[1 4C]leucine («Amersham») diluted with non-labelled
amino acid to 68420 cpm/nmol, 4 mg/ml unfrac-
tionated E. coli tRNA and 0.001—1 mg/ml of protein
depending upon the purity of the enzyme. The
reaction was conducted at 55 °С for 1 min and the
samples were placed on GF/A filters. The filters were
washed three times in 5 % trichloroacetic acid,
ethanol, 50:50 (vol/vol.) ethanol/ether and the ra
dioactivity was measured by liquid scintillation co
unting.
The Km values for ATP, leucine and tRNA U u
were determined at 65 °С in the standard mixture
containing 10 jug/ml of LeuRSTT and varying con
centration of ATP (10—300 / Ш ) , leucine (0.5—
50 juM) or tRNA (0.05—2.5 pM) respectively (using
ENZFITTER programme).
The molecular mass determination. The mo
lecular mass of the enzyme was determined by gel
filtration on a column (1 .1x72 cm) of Sephadex
G-200 and by PAGE under native and denaturing
conditions. Under native conditions it was conducted
in 5 % and 6 % polyacrylamide gels. Under de
naturing conditions it was conducted in 12.5 %
polyacrylamide gel in the presence of 0.1 % SDS as
described by Laemmli [9].
Measurements of thermal stability of LeuRS. The
heat stability of the aminoacylation activity of
LeuRSTT was analyzed by incubating 100 /Л of the
enzyme solution (1.5 mg/ml) in 100 mM Na-Hepes,
pH 7.3 containing 15 mM MgCl2 with or without the
small substrates at the indicated concentration. At
various times, 10-/Л aliquots were removed, diluted in
cold enzyme dilution buffer containing 100 mM Na-
Hepes, pH 7.3, 2 mM DTT, 10 % glycerol, 15 mM
MgCl2 and 5 m g / m l BSA. T h e r ema in ing
aminoacylation activity was determined by initial rate
measurements as described above.
Crystallization procedure. Crystallization trials
were conducted at different constant temperatures (6,
12, and 20 °С) using the hanging-drop vapor-dif
fusion method. The drops of 4—10 jul were allowed to
equilibrate against 0.8 ml reservoir solution. Con
ditions such as the nature of the precipitant, the
composition and pH of the buffer and the con
centration of the protein were varied.
Results and Discussion. LeuRSTT was purified
from both HB8 and HB27 strains to homogeneity
using a five-step purification procedure (Table 1).
The final yield was approximately 20 mg of the pure
enzyme from 520 g cells with a specific activity of
2212 U/mg (1 U of the enzyme catalyzes the
formation of 1 nmol leucyl-tRNA/min at 65 °С). Gel
filtration gives M r of 100 kDa. PAGE under native
conditions reveals an apparent M r of 102 kDa.
SDS/PAGE shows a single band corresponding to a
polypeptide chain of about 101 kDa. The kinetic
constants of the aminoacylation reaction catalyzed by
217
YAREMCHUK A. D . E T AL.
Table I
Purification of leucyl-tRNA synthetase from T. thermophilus HB8
•Obtained from 520 g cells.
Table 2
Kinetic constants of aminoacylation reaction catalyzed by LeuRS
from T. thermophilus at 65 °С
LeuRSTT for ATP, leucine and tRNA U u are reported
in Table 2. There is no significant difference in Km
and кш values of LeuRSTT for all three substrates
compared with those of LeuRS from E. coli [10]. The
optimal temperature for the tRNA aminoacylation
reaction catalyzed by LeuRSTT is about 70 °С for
tRNA U u from T. thermophilus and about 65 °С for
tRNA U u from E. coli (Fig. 1). The thermal stability
of the enzyme was investigated in the range from 50
to 90 °С. The enzyme is remarkably thermostable. It
retains 100 % of its ability to aminoacylate tRNA
after 1 h of incubation at 85 °С (data not shown) and
about 97 % at 88 °С. ATP and leucine alone do not
affect the thermal inactivation of the enzyme at 88 °С.
Surprisingly, when ATP and leucine or ATP, leucine
and inorganic pyrophosphate are present together
significant inactivation of LeuRS occurs after 1 h at
88 °С (Fig. 2). In contrast, the protection of GlyRS
from thermoinactivation by ATP, glycine or gly-
cyladenylate was shown for T. thermophilus GlyRS
[11], ATP or tyrosyladenylate for TyrRS, histidine
or histidyladenylate for HisRS and prolyladenylate
for ProRS from T. thermophilus (Yaremchuk et al.,
unpublished results). In all these cases the protection
of the aaRS from thermal inactivation by substrates
correlates with conformational changes of the enzyme
promoted by substrates binding ([11], Cusack S.,
Yaremchuk A. and Tukalo M., unpublished results).
To study the mechanism of amino acid re
cognition and activation and the specific recognition of
cognate tRNA by T. thermophilus LeuRS we tried to
crystallize this enzyme alone and in complex with the
substrates. Initial screening of crystallization con
ditions was conducted using a sparse matrix sampling
reagents from Hampton Research (Crystal screen 1
and 2, Grid screens including ammonium sulfate,
Fig. 1. Dependence of the rate of tRNA aminoacylation catalyzed by
LeuRSTT on temperature: 1 — T thermophilus t R N A U u ; 2 — E.
coli t R N A U u
218
LEUCYL-tRNA S Y N T H E T A S E FROM T H E R M U S THERMOPHILUS
Relative activity, %
0 20 40 Time, min
Fig. 2. Thermal stability of LeuRSTT in the absence and in the
presence of small ligands. The enzyme (1.5 mg/ml) was incubated
at 88 °С in 100 mM Na-Hepes, pH 7.3 , and 15 mM MgCl 2 either in
the absence (7) or in the presence (2) of 1 mM ATP and 2 mM
leucine and in the presence (3) of 1 mM ATP, 2 mM leucine and 1
mM PP,. The remaining activity was measured after 0—60 min
incubation at 88 °С
polyethylene glycol 6000, sodium chloride, 2-methyl-
2,4-pentanediol and Crystal screen cryo formulation)
without any success. Precipitants alone, such as
ammonium sulfate, PEG, sodium formate, sodium
citrate, sodium chloride and MPD at different con
centrations over a broad range of pH and protein
concentration did not yield crystals suitable for X-ray
structure determination. Extremely thin needle-like
crystals were obtained from 48 % ammonium sulfate
solution at pH 7.5—8.3 at 6 °С and were not of
suitable dimensions for diffraction experiments. Not
well-shaped crystals (Fig. 3, a) and diffracting to 9 A
resolution were obtained at 20 °С from 38—40 %
ammonium sulfate solution at pH 7.5 in the presence
of 5 % glycerol after two—three weeks and are very
poorly reproducible. SDS/PAGE shows degradation of
the enzyme after one week at 20 °С and even crystals
contain two polypeptide chains with molecular mass of
about 100 and 96 kDa (data not shown). Better-
quality crystals of LeuRSTT were obtained under the
following conditions: to, prevent protein degradation
10/Л drops containing 8—10 mg/ml LeuRS in 40 mM
Tris-HCl, pH 7.5, 2 mM DTT, 10 mM MgCl2, 1 mM
NaN 3, and 15 % ammonium sulfate were equilibrated
for 12—14 hours at 6 °С against 800 ml reservoir
solution containing 54—58 % ammonium sulfate in
100 mM Tris-HCl, pH 7.5. After rapid precipitation
of the macromolecules the drops were transferred to
12 °С and ammonium sulfate concentration was
Fig, 3. Crystals of LeuRSTT: a — not well-shaped crystals and
diffracting to 9 A resolution; b — crystalls obtained by crystallization
from precipitate and diffracting to 3 .5 A resolution
decreased to 40—42 %. Under these conditions
well-shaped crystals appeared in two days and grew
from precipitate rapidly over a 4—5 days period to a
maximum of 0,8 mm in the longest dimension (Fig.
3, b). After stabilization for one week against 48 %
saturated ammonium sulfate in 100 mM Tris-HCl, pH
7.5 crystals were mounted in quartz capillaries with a
small amount of reservoir solution for X-ray dif
fraction measurements. These crystals diffract to
3.5 A resolution. The protein from washed and solu-
bilized crystals as well as the protein used for
crystallization was verified as full-length LeuRSTT by
SDS/PAGE and enzymatic ability to aminoacylate
tRNA. X-ray diffraction analysis of these crystals is
under way. Small crystals of LeuRSTT in complex
219
YAREMCHUK A. D . ET AL.
with tRNA U u were obtained using ammonium sulfate
as a precipitant but they were not of suitable dimen
sions for X-ray analysis. The gene for LeuRSTT has
been recently cloned, sequenced end expressed in E.
coli (Tukalo et aL, unpublished results) and the work
is under way to obtain high diffracting crystals of
recombinant LeuRSTT and its complexes with sub
strates.
Acknowledgements. The research was supported
in part by an International Research Scholar's award
from the Howard Hughes Medical Institute.
Г. Д. Яре мчу к, О. I. Гудзера, С. П. Егорова, Д. I. Рожко,
I. А. Крикливий, М. А. Тукало
Лейцил-тРНК синтетаза із Thermus thermophilus. Очищення і
деякі властивості кристалів ферменту
Резюме
Лейцил-тРНК синтетазу із Thermus thermophilus (ЛейРСТТ)
виділено в гомогенному стані з використанням п'яти стадій
очищення. Фермент охарактеризовано та отримано його
кристали. Визначено молекулярну масу нашивного і денатуро
ваного білка. Встановлено, що ЛейРСТТ являє собою мономер
з молекулярною масою 101 кДа. Отриманому ферментові
притаманна значна термостабільність і він зберігає 97 %
аміноацилюючої активності після інкубації протягом 1 год
при температурі 88 ° С Кристали ЛейРСТТ одержано мето
дом дифузії парів із використанням як осаджувана розчину
сульфату амонію.
А. Д. Яремчук, О. И. Гудзера, С. П. Егорова, Д. И. Рожко,
И. А. Крикливый, М. А. Тукало
Лейцил-тРНК синтетаза из Thermus thermophilus. Очистка и
некоторые свойства кристаллов фермента
Резюме
Лейцил-тРНК синтетаза из Thermus thermophilus (ЛейРСТТ)
выделена в гомогенном состоянии с использованием пяти
стадий очистки. Фермент охарактеризован и получены его
кристаллы. Определена молекулярная масса нашивного и дена
турированного белка. Установлено, что ЛейРСТТ представ
ляет собой мономер с молекулярной массой 101 кДа. Получен
ный фермент обладает значительной термостабильностью и
сохраняет 97 % аминоацилирующей активности после инкуба
ции в течение 1 ч при температуре 88 °С. Кристаллы
ЛейРСТТ получены методом диффузии паров с использованием
в качестве осадителя раствора сульфата аммония.
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УДК 577.112.088.3
Надійшла до редакції 27.01.2000
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| id | nasplib_isofts_kiev_ua-123456789-155227 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0233-7657 |
| language | English |
| last_indexed | 2025-12-07T17:48:25Z |
| publishDate | 2001 |
| publisher | Інститут молекулярної біології і генетики НАН України |
| record_format | dspace |
| spelling | Yaremchuk, A.D. Gudzera, O.I. Egorova, S.P. Rozhko, D.I. Kriklivy, I.A. Tukalo, M.A. 2019-06-16T11:53:38Z 2019-06-16T11:53:38Z 2001 Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme / A.D. Yaremchuk, O.I. Gudzera, S.P. Egorova, D.I. Rozhko, I.A. Kriklivy, M.A. Tukalo // Біополімери і клітина. — 2001. — Т. 17, № 3. — С. 216-220. — Бібліогр.: 11 назв. — англ. 0233-7657 DOI:http://dx.doi.org/10.7124/bc.0005AD https://nasplib.isofts.kiev.ua/handle/123456789/155227 577.112.088.3 Leucyl-tRNA synthetase from Thermus thermophilus (LeuRSTT) was purified to homogeneity using a five-step purification procedure. The enzyme was characterized and crystallized. Molecular mass determinations of the native and denatured proteins indicate monomeric structure of LeuRSTT with the molecular mass of about 101 kDa. The protein obtained is remarkably thermostable and retains 97 % of its initial aminoacylation activity after 1 hour of incubation at 88 °C. Crystals of LeuRSTT were obtained from ammonium sulfate solution by the vapour diffusion techniques. The crystals quality was improved by crystallization from the precipitate. Лейцил-тРНК синтетаза из Thermus thermophilus (ЛейРСТТ) выделена в гомогенном состоянии с использованием пяти стадий очистки. Фермент охарактеризован и получены его кристаллы. Определена молекулярная масса нашивного и денатурированного белка. Установлено, что ЛейРСТТ представляет собой мономер с молекулярной массой 101 кДа. Полученный фермент обладает значительной термостабильностью и сохраняет 97 % аминоацилирующей активности после инкубации в течение 1 ч при температуре 88 °С. Кристаллы ЛейРСТТ получены методом диффузии паров с использованием в качестве осадителя раствора сульфата аммония Лейцил-тРНК синтетазу із Thermus thermophilus (ЛейРСТТ) виділено в гомогенному стані з використанням п'яти стадій очищення. Фермент охарактеризовано та отримано його кристали. Визначено молекулярну масу нативного і денатурованого білка. Встановлено, що ЛейРСТТ являє собою мономер з молекулярною масою 101 кДа. Отриманому ферментові притаманна значна термостабільність і він зберігає 97 % аміноацилюючої активності після інкубації протягом 1 год при температурі 88 ° С Кристали ЛейРСТТ одержано методом дифузії парів із використанням як осаджувана розчину сульфату амонію. en Інститут молекулярної біології і генетики НАН України Біополімери і клітина Структура та функції біополімерів Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme Лейцил-тРНК синтетаза із Thermus thermophilus. Очищення і деякі властивості кристалів ферменту Лейцил-тРНК синтетаза из Thermus thermophilus. Очистка и некоторые свойства кристаллов фермента Article published earlier |
| spellingShingle | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme Yaremchuk, A.D. Gudzera, O.I. Egorova, S.P. Rozhko, D.I. Kriklivy, I.A. Tukalo, M.A. Структура та функції біополімерів |
| title | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme |
| title_alt | Лейцил-тРНК синтетаза із Thermus thermophilus. Очищення і деякі властивості кристалів ферменту Лейцил-тРНК синтетаза из Thermus thermophilus. Очистка и некоторые свойства кристаллов фермента |
| title_full | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme |
| title_fullStr | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme |
| title_full_unstemmed | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme |
| title_short | Leucyl-tRNA synthetase from Thermus thermophilus. Purification and some properties of the crystallizing enzyme |
| title_sort | leucyl-trna synthetase from thermus thermophilus. purification and some properties of the crystallizing enzyme |
| topic | Структура та функції біополімерів |
| topic_facet | Структура та функції біополімерів |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/155227 |
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