PDB-8vu0: Co-crystal structure of Aquifex aeolicus Trbp111 in complex with ...

ID or keywords:

Entry

Database: PDB / ID: 8vu0

Title

Co-crystal structure of Aquifex aeolicus Trbp111 in complex with E. coli tRNA-Ile

Components

Methionyl-tRNA synthetase beta subunit
RNA (76-MER)

Keywords

RNA BINDING PROTEIN/RNA / OB fold / EMAPII-like domain / tRNA-binding / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA complex

Function / homology

Function and homology information

methionine-tRNA ligase activity / methionyl-tRNA aminoacylation / tRNA binding / ATP binding
Similarity search - Function

Biological species

Aquifex aeolicus (bacteria)

Escherichia coli (E. coli)

Method

X-RAY DIFFRACTION /

SYNCHROTRON /

MOLECULAR REPLACEMENT / Resolution: 2.64 Å

Authors

Umuhire Juru, A. / Zhang, J.

Funding support

United States, 1items

Citation

Journal: Nat Commun / Year: 2024
Title: Structural basis of tRNA recognition by the widespread OB fold.
Authors: Umuhire Juru, A. / Ghirlando, R. / Zhang, J.

History

Deposition	Jan 27, 2024	Deposition site: RCSB / Processing site: RCSB
Revision 1.0	Aug 7, 2024	Provider: repository / Type: Initial release

Structure viewer	Molecule: MolmilJmol/JSmol

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Download

PDBx/mmCIF format	8vu0.cif.gz	318.5 KB	Display	PDBx/mmCIF format
PDB format	pdb8vu0.ent.gz	211.8 KB	Display	PDB format
PDBx/mmJSON format	8vu0.json.gz		Tree view	PDBx/mmJSON format
Others	Other downloads

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Validation report

Summary document	8vu0_validation.pdf.gz	457.1 KB	Display	wwPDB validaton report
Full document	8vu0_full_validation.pdf.gz	460.9 KB	Display
Data in XML	8vu0_validation.xml.gz	15.9 KB	Display
Data in CIF	8vu0_validation.cif.gz	23.7 KB	Display
Arichive directory	https://data.pdbj.org/pub/pdb/validation_reports/vu/8vu0 ftp://data.pdbj.org/pub/pdb/validation_reports/vu/8vu0	HTTPS FTP

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Related structure data

Related structure data	8vtzC C: citing same article (ref.)
Similar structure data	Similarity search - Function & homologyF&H Search Similarity search - FunctionF&H Search Similarity search - HomologyF&H Search

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Links

PDB pages	PDBj / wwPDB / NCBI
Related items in Molecule of the Month	#121 - Jan 2010 70S Ribosomes similarity (1) #16 - Apr 2001 Aminoacyl-tRNA Synthetases similarity (4) #130 - Oct 2010 Riboswitches similarity (1) #157 - Jan 2013 Transfer-Messenger RNA similarity (1) #181 - Jan 2015 Cascade and CRISPR similarity (1) #229 - Jan 2019 Fluorescent RNA Aptamers similarity (1) #253 - Jan 2021 Expressome similarity (1) #260 - Aug 2021 Ribonuclease P similarity (1) #146 - Feb 2012 Aminoglycoside Antibiotics similarity (1) #15 - Mar 2001 Transfer RNA similarity (1) #249 - Sep 2020 SARS-CoV-2 RNA-dependent RNA Polymerase similarity (1) #10 - Oct 2000 Ribosome similarity (1) #210 - Jun 2017 Adenine Riboswitch in Action similarity (1) #87 - Mar 2007 Zinc Fingers similarity (1) #105 - Sep 2008 Ribonuclease A similarity (1) #226 - Oct 2018 Aminoglycoside Antibiotics and Resistance similarity (1) #245 - May 2020 Spliceosomes similarity (1) #262 - Oct 2021 Fifty Years of Open Access to PDB Structures similarity (1) #65 - May 2005 Self-splicing RNA similarity (1) #98 - Feb 2008 Small Interfering RNA similarity (1) #104 - Aug 2008 Selenocysteine Synthase similarity (1) #106 - Oct 2008 Poly(A) Polymerase similarity (1) #143 - Nov 2011 Toll-like Receptors similarity (1) #161 - May 2013 Ricin similarity (1) #227 - Nov 2018 Telomerase similarity (1) #230 - Feb 2019 Initiation Factor eIF4E similarity (1) #266 - Feb 2022 Oligosaccharyltransferase similarity (1) #81 - Sep 2006 Elongation Factors similarity (1) #89 - May 2007 Aconitase and Iron Regulatory Protein 1 similarity (1) #241 - Jan 2020 Twenty Years of Molecules similarity (1)

PDB pages

PDBj /

wwPDB /

NCBI

Related items in Molecule of the Month

#121 - Jan 2010
70S Ribosomes
similarity (1)
#16 - Apr 2001
Aminoacyl-tRNA Synthetases
similarity (4)
#130 - Oct 2010
Riboswitches
similarity (1)
#157 - Jan 2013
Transfer-Messenger RNA
similarity (1)
#181 - Jan 2015
Cascade and CRISPR
similarity (1)
#229 - Jan 2019
Fluorescent RNA Aptamers
similarity (1)
#253 - Jan 2021
Expressome
similarity (1)
#260 - Aug 2021
Ribonuclease P
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#146 - Feb 2012
Aminoglycoside Antibiotics
similarity (1)
#15 - Mar 2001
Transfer RNA
similarity (1)
#249 - Sep 2020
SARS-CoV-2 RNA-dependent RNA Polymerase
similarity (1)
#10 - Oct 2000
Ribosome
similarity (1)
#210 - Jun 2017
Adenine Riboswitch in Action
similarity (1)
#87 - Mar 2007
Zinc Fingers
similarity (1)
#105 - Sep 2008
Ribonuclease A
similarity (1)
#226 - Oct 2018
Aminoglycoside Antibiotics and Resistance
similarity (1)
#245 - May 2020
Spliceosomes
similarity (1)
#262 - Oct 2021
Fifty Years of Open Access to PDB Structures
similarity (1)
#65 - May 2005
Self-splicing RNA
similarity (1)
#98 - Feb 2008
Small Interfering RNA
similarity (1)
#104 - Aug 2008
Selenocysteine Synthase
similarity (1)
#106 - Oct 2008
Poly(A) Polymerase
similarity (1)
#143 - Nov 2011
Toll-like Receptors
similarity (1)
#161 - May 2013
Ricin
similarity (1)
#227 - Nov 2018
Telomerase
similarity (1)
#230 - Feb 2019
Initiation Factor eIF4E
similarity (1)
#266 - Feb 2022
Oligosaccharyltransferase
similarity (1)
#81 - Sep 2006
Elongation Factors
similarity (1)
#89 - May 2007
Aconitase and Iron Regulatory Protein 1
similarity (1)
#241 - Jan 2020
Twenty Years of Molecules
similarity (1)

Deposited unit

C: RNA (76-MER)

D: RNA (76-MER)

B: Methionyl-tRNA synthetase beta subunit

A: Methionyl-tRNA synthetase beta subunit

73.7 kDa, 4 polymers
Search similar-shape structures of this assembly by Omokage search (details)

1

Idetical with deposited unit
defined by author
Evidence: equilibrium centrifugation

Unit cell

Length a, b, c (Å)	190.495, 55.028, 87.160
Angle α, β, γ (deg.)	90.000, 94.420, 90.000
Int Tables number	5
Space group name H-M	C121
Space group name Hall	C2y

Noncrystallographic symmetry (NCS)

NCS domain:

ID	Ens-ID	Details
d_1	ens_1	chain "A"
d_2	ens_1	chain "B"
d_1	ens_2	(chain "C" and (resid 1 through 15 or resid 18 through 76))
d_2	ens_2	(chain "D" and (resid 1 through 15 or resid 18 through 76))

NCS domain segments:

Component-ID: 1

Dom-ID	Ens-ID	Beg auth comp-ID	Beg label comp-ID	End auth comp-ID	End label comp-ID	Auth asym-ID	Label asym-ID	Auth seq-ID	Label seq-ID
d_1	ens_1	ALA	ALA	SER	SER	A	D	5 - 111	7 - 113
d_2	ens_1	ALA	ALA	SER	SER	B	C	5 - 111	7 - 113
d_1	ens_2	G	G	A	A	C	A	1 - 9	1 - 9
d_2	ens_2	G	G	A	A	D	B	1 - 9	1 - 9

NCS ensembles :

ID
ens_1
ens_2

#1: RNA chain	RNA (76-MER) Mass: 24604.627 Da / Num. of mol.: 2 / Source method: obtained synthetically / Source: (synth.) Escherichia coli (E. coli)
#2: Protein	Methionyl-tRNA synthetase beta subunit Mass: 12266.301 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Aquifex aeolicus (bacteria) / Gene: metG', aq_422 / Production host: Escherichia coli (E. coli) / References: UniProt: O66738
#3: Water	ChemComp-HOH / water Mass: 18.015 Da / Num. of mol.: 201 / Source method: isolated from a natural source / Formula: H₂O

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Experiment

Experiment	Method: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

Crystal	Density Matthews: 3.09 Å³/Da / Density % sol: 60.17 %
Crystal grow	Temperature: 294.15 K / Method: vapor diffusion, hanging drop / Details: 0.2 M ammonium phosphate dibasic, 23% PEG 3350

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Data collection

Diffraction	Mean temperature: 100 K / Serial crystal experiment: N
Diffraction source	Source: SYNCHROTRON / Site: APS / Beamline: 22-ID / Wavelength: 1 Å
Detector	Type: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Dec 16, 2022
Radiation	Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength	Wavelength: 1 Å / Relative weight: 1
Reflection	Resolution: 2.64→43.09 Å / Num. obs: 26578 / % possible obs: 98.79 % / Redundancy: 12.6 % / B_iso Wilson estimate: 59.56 Å² / CC_1/2: 0.977 / R_pim(I) all: 0.0438 / Net I/σ(I): 9.76
Reflection shell	Resolution: 2.64→2.73 Å / Mean I/σ(I) obs: 0.87 / Num. unique obs: 2645 / CC_1/2: 0.909 / R_pim(I) all: 0.197

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Processing

Software

Name	Version	Classification
DIALS		data scaling
DIALS		data reduction
PHASER		phasing
PHENIX	1.20.1_4487	refinement

Refinement

Method to determine structure:

MOLECULAR REPLACEMENT / Resolution: 2.64→43.09 Å / SU ML: 0.4524 / Cross valid method: FREE R-VALUE / σ(F): 1.33 / Phase error: 24.9231
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2

	R_factor	Num. reflection	% reflection
R_free	0.2217	1319	4.97 %
R_work	0.1825	25211	-
obs	0.1845	26530	98.82 %

Solvent computation

Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL

Displacement parameters

B_iso mean: 57.49 Å²

Refinement step

Cycle: LAST / Resolution: 2.64→43.09 Å

	Protein	Nucleic acid	Ligand	Solvent	Total
Num. atoms	1628	3262	0	201	5091

Refine LS restraints

Refine-ID	Type	Dev ideal	Number
X-RAY DIFFRACTION	f_bond_d	0.0031	5288
X-RAY DIFFRACTION	f_angle_d	0.739	7894
X-RAY DIFFRACTION	f_chiral_restr	0.0426	1032
X-RAY DIFFRACTION	f_plane_restr	0.0048	430
X-RAY DIFFRACTION	f_dihedral_angle_d	13.5684	2046

Refine LS restraints NCS

Ens-ID	Dom-ID	Asym-ID	Auth asym-ID	Refine-ID	Type	Rms dev position (Å)
ens_1	d_2	D	A	X-RAY DIFFRACTION	Torsion NCS	0.668569009287
ens_2	d_2	A	C	X-RAY DIFFRACTION	Torsion NCS	0.805739687209

LS refinement shell

Resolution (Å)	R_factor R_free	Num. reflection R_free	R_factor R_work	Num. reflection R_work	Refine-ID	% reflection obs (%)
2.64-2.75	0.3978	151	0.3792	2780	X-RAY DIFFRACTION	99.29
2.75-2.87	0.3877	155	0.312	2781	X-RAY DIFFRACTION	99.16
2.87-3.02	0.3455	137	0.2515	2771	X-RAY DIFFRACTION	98.88
3.02-3.21	0.2898	145	0.2347	2755	X-RAY DIFFRACTION	97.84
3.21-3.46	0.2636	133	0.2135	2821	X-RAY DIFFRACTION	99.56
3.46-3.81	0.2345	148	0.1683	2823	X-RAY DIFFRACTION	99.5
3.81-4.36	0.1869	141	0.153	2801	X-RAY DIFFRACTION	99.16
4.36-5.49	0.1706	148	0.1347	2818	X-RAY DIFFRACTION	97.98
5.49-43.09	0.1568	161	0.1486	2861	X-RAY DIFFRACTION	98.12

Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

ID	L11 (°²)	L12 (°²)	L13 (°²)	L22 (°²)	L23 (°²)	L33 (°²)	S11 (Å °)	S12 (Å °)	S13 (Å °)	S21 (Å °)	S22 (Å °)	S23 (Å °)	S31 (Å °)	S32 (Å °)	S33 (Å °)	T11 (Å²)	T12 (Å²)	T13 (Å²)	T22 (Å²)	T23 (Å²)	T33 (Å²)	Origin x (Å)	Origin y (Å)	Origin z (Å)
1	7.46506630477	0.0356711864404	0.861266037928	8.86737221059	0.541269938099	2.82850416474	-0.384476139741	-0.00553727843196	-0.0918952700299	-0.106728389141	-0.165132511581	-0.409422841277	0.508972869118	0.44121291806	0.481216843445	0.253964326089	0.0233897994522	0.0106037313361	0.284146389839	-0.000231334725985	0.404165250659	45.9967293816	-40.2811479142	-21.0922837625
2	8.87826871497	4.71499530761	-4.36043115212	7.25740298119	-6.07950586115	6.80407885163	-0.411057554781	0.377591672047	0.383136307799	-0.516303518442	1.18533869173	1.02787484649	-0.317613379116	-0.663052238133	-0.342486721264	0.374786277236	-0.0142075667893	0.0229352641432	0.427649368263	-0.0862963272814	0.456237834446	27.9526215471	-42.3961413271	-24.689039405
3	6.82060210013	4.46955227421	0.467412888507	7.94236290481	4.08029985784	5.23132429248	0.0739311561697	0.202280310251	0.246220554389	0.378345158133	0.257119687855	0.0979082747823	-0.43555458413	0.0199866818593	-0.282010062592	0.435638443067	0.0192306283697	0.015452179734	0.307881410677	0.0809374930718	0.385286753646	44.2256547453	-24.4891106493	-19.9824528351
4	2.01773187051	-1.0280770357	-1.52073400084	4.59587725823	-3.73290924263	4.33780552536	-0.306753211009	-1.50921079029	0.743447165336	1.63544104608	-0.503064270371	-0.614829694607	-0.861457442607	-0.766300137689	0.751694493117	0.495480829532	-0.0552808401653	-0.0983476069333	0.553469494934	-0.0521828597866	0.396609723956	40.6130864679	-20.1816675999	-3.73933068063
5	2.39330565893	-1.56766239086	-2.40858289527	3.10145091085	3.01594228106	3.42767538466	-0.135992386679	-0.288418828109	-0.0416995730965	-0.290390269656	-0.310875366219	0.160904687936	-1.33618060614	-0.594086281383	0.645335151881	0.45908959262	0.0349810360713	-0.099580604579	0.444120470474	0.0173443180024	0.500314992722	42.6389591296	-21.3543156907	-17.8988003203
6	4.6141061187	-4.37893746515	-4.95129279608	8.55792843851	5.51028842054	5.63228057056	0.474394189743	0.569763552052	0.534765203156	-0.994602317619	-0.577947709777	0.649756953858	-2.25094246712	-0.643840223803	0.0830806415517	0.558947692516	0.0686704308472	-0.0850671266714	0.600317131712	0.0261882923521	0.604767558222	38.806864041	-22.4298613207	-23.8094679748
7	0.924617899156	-0.322711491542	-1.59935877349	2.98819332071	1.10175282432	3.77327946627	-0.149058476058	0.0317813175955	0.46494709194	-0.0384984961547	-0.0507095934205	0.102882114245	-0.179469339971	0.379858326954	0.122306927026	0.270990394171	0.0114559450621	-0.0724357009954	0.30899957369	-0.035171811657	0.522410321061	43.7607675272	-27.8678325106	-15.4350504366
8	4.51064639566	5.17361089862	4.06463798832	9.39080578304	5.19458161629	3.88987196844	-0.0554438714317	0.264974185711	0.666118933318	-0.294967795483	0.0171383576016	0.393419772775	-0.72412742747	0.0104593859192	0.168166454647	0.361283574874	0.0895415576712	-0.0213509606567	0.483306103357	-0.0956439575548	0.506397119432	32.2895020286	-20.198393697	-13.5253597225
9	6.92739880244	2.30030265599	-2.28607091145	6.32999428377	-4.92287731104	3.85961982932	-0.384294835298	-0.628720723869	-0.562892149966	0.631888449526	-0.894602522885	0.404129252952	1.04424884657	-0.0154332604915	1.30012836499	0.625293241104	0.0958156041012	0.1491693622	0.295733315047	-0.0988604350229	0.538377285153	39.6841959346	-34.1259914079	-14.1257737057
10	8.1096573324	0.43140359336	-1.5940213907	8.47428346648	0.437499337899	9.07954163578	-0.100657714073	0.260015829456	0.139269162743	-0.0587925460092	0.119140122296	0.002786431246	0.0218796197872	0.571756821466	-0.051795575267	0.173983376381	0.0268541361972	-0.0283448755249	0.305795001959	-0.0285661096034	0.392913827033	46.1249692034	-36.2666904667	-21.3033325598
11	2.3249750327	-0.454557675912	1.6732251772	3.47297622964	-0.00718534625261	1.51323288775	-0.104566415756	-0.112987067706	-0.319580259369	0.104911393717	0.195953270805	0.136723252003	0.14590506576	-0.214185417238	-0.0202530101558	0.359188396799	0.0418602557699	0.0712240279713	0.418343137706	-0.0526407117618	0.479004356548	17.7863348209	16.5911066983	4.70100838789
12	0.652261862167	0.129306004662	0.703673540115	-0.119574620335	0.326318010727	-0.498242778965	-0.431145893227	-1.20917062572	0.952498268184	0.118424968214	0.289727981002	-0.130936552762	0.027674253393	-0.356520233603	0.196436204912	0.41916813572	0.118609672421	0.00872378511384	0.904979070804	-0.135526296858	0.880714579703	1.71810386298	26.47659727	10.8801281829
13	0.187366373215	0.268915662734	-0.563124395194	8.27302194271	0.638578361743	0.342358785298	0.120614526437	0.0134062046998	-0.211676232192	0.228277633579	-0.305266038153	0.0216672825709	0.0871411846025	-0.0500692179981	0.172154413556	0.383382842347	-0.0218051679174	-0.0336777256782	0.487806509088	-0.0701831066808	0.564038848314	31.7135411036	4.75352478487	5.12382509957
14	2.73901788912	-0.0223973709351	-1.72616566882	2.03901778256	-0.458986793029	2.0379284246	-0.237675195438	0.0311510017917	0.0299145730086	-0.218469721151	0.381494088384	0.237411153623	-0.235245374377	-0.492419709817	0.00377569789556	0.41753581269	-0.053377228557	-0.08464740706	0.442658511673	-0.0360745158877	0.541540589385	21.9309375539	-93.8196152329	-47.9258055426
15	1.5531912283	-0.180562894526	-1.01547926107	-0.0886716976314	0.506357331047	-0.402273594477	-0.417060864558	1.29920213822	-1.15092922165	-0.15657649653	0.224971190626	-0.0822902509808	-0.120404518347	-0.319083639526	0.189966347626	0.455457974921	-0.155472806619	0.0584192368147	0.898012997385	-0.131081095509	0.846548166574	5.35082767431	-103.642139707	-54.3972303122
16	0.30364380546	0.669721702041	0.811726469832	7.50809181603	0.0163750875954	0.207980744076	-0.00431484941084	0.0906363091965	0.219086620345	-0.201790348396	-0.228339102908	-0.269577256909	-0.161594867582	0.0210234937863	0.198741475941	0.483577618766	-0.0105894416145	0.127242627398	0.465362027149	-0.0478839764043	0.556521491746	35.9768349045	-83.1161219909	-48.535207242
17	3.46978699848	0.935202170603	-0.262804194638	4.32902296534	-6.24220554291	9.4018557473	-0.402306645213	-0.458747266633	-0.00698743677659	-0.554432682793	0.654647913221	0.558303438745	0.479487169089	-1.31661410846	-0.0584958456459	0.373588346554	3.93077241387E-5	0.0284272908033	0.483894185356	-0.0782259723483	0.555409948302	27.412621749	-36.1911101787	-18.6183339846
18	2.74106113109	0.366589962187	1.97954862595	4.42619241581	1.77249088976	7.20662127176	0.0843841000462	0.395974789722	-0.597237411846	0.109482657816	0.309847613688	-0.454512702641	1.26386270699	0.770691476297	-0.458346033992	0.348346340963	0.0590725484595	-0.0373812713649	0.307144905822	-0.0692003743962	0.512232653868	45.208827198	-54.0440824757	-27.8045475482
19	1.40180930801	0.538772885737	-0.199288211967	1.80519990106	1.62856498105	6.65818498942	-0.0574215153756	-0.0965321228517	-0.131552371768	0.216253889161	-0.0284547896598	-0.276583605014	0.514447985604	0.217238639514	0.0910096018665	0.427310712324	0.0634175030557	0.0318516707574	0.404517530737	-0.0648844998183	0.682843998292	45.8377037503	-51.5148209191	-24.0580598077
20	4.70328550257	-4.87040272524	-4.50647652259	5.24194821904	4.8146719728	4.79923958224	0.0604249408124	0.182305418533	-0.926219331868	0.1025790059	-0.507546485641	1.06972155879	0.448228165638	-0.903357511128	0.669582244719	0.547217204765	-0.0155717802875	0.0675942483043	0.417496927004	-0.0545596405238	0.586246666038	34.5000901541	-53.8495532528	-26.5928881898

Refinement TLS group

Refine-ID: X-RAY DIFFRACTION

ID	Refine TLS-ID	Selection details	Auth asym-ID	Label asym-ID	Auth seq-ID	Label seq-ID
1	1	chain 'B' and (resid 88 through 111 )	B	C	88 - 111	84 - 107
2	2	chain 'A' and (resid 5 through 16 )	A	D	5 - 16	1 - 12
3	3	chain 'A' and (resid 17 through 27 )	A	D	17 - 27	13 - 23
4	4	chain 'A' and (resid 28 through 34 )	A	D	28 - 34	24 - 30
5	5	chain 'A' and (resid 35 through 40 )	A	D	35 - 40	31 - 36
6	6	chain 'A' and (resid 41 through 50 )	A	D	41 - 50	37 - 46
7	7	chain 'A' and (resid 51 through 74 )	A	D	51 - 74	47 - 70
8	8	chain 'A' and (resid 75 through 84 )	A	D	75 - 84	71 - 80
9	9	chain 'A' and (resid 85 through 90 )	A	D	85 - 90	81 - 86
10	10	chain 'A' and (resid 91 through 111 )	A	D	91 - 111	87 - 107
11	11	chain 'C' and (resid 1 through 30 )	C	A	1 - 30
12	12	chain 'C' and (resid 31 through 50 )	C	A	31 - 50
13	13	chain 'C' and (resid 51 through 76 )	C	A	51 - 76
14	14	chain 'D' and (resid 1 through 30 )	D	B	1 - 30
15	15	chain 'D' and (resid 31 through 50 )	D	B	31 - 50
16	16	chain 'D' and (resid 51 through 76 )	D	B	51 - 76
17	17	chain 'B' and (resid 5 through 16 )	B	C	5 - 16	1 - 12
18	18	chain 'B' and (resid 17 through 34 )	B	C	17 - 34	13 - 30
19	19	chain 'B' and (resid 35 through 69 )	B	C	35 - 69	31 - 65
20	20	chain 'B' and (resid 70 through 87 )	B	C	70 - 87	66 - 83

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