Structural and hydration changes in the active site gorge of phosporhylated butyrylcholinesterase accompanying the aging process.


Abstract

Wild-type (wt) butyrylcholinesterase (BuChE) and the E197D and D70G mutants were inhibited by diisopropylfluorophosphate (DFP) or soman under standard conditions of pH, temperature and pressure. The effect of hydrostatic and osmotic pressures on the aging process of DFP-phosphorylated enzymes (diisopropylphosphoryl-BuChE (DIP-BuChE)) was investigated. Hydrostatic pressure strongly increased the rate of aging of wt enzyme. The activation volumes (deltaV*) for the dealkylation reaction was -150 ml/mol for DIP-wtBuChE. On the other hand, pressure had little effect on the aging of the DIP-E197D mutant and no effect on the DIP-D70G mutant, indicating that the transition state of the aging reaction (dealkylation of an isoproxy chain) was associated with an extended conformation/hydration change in wtBuChE but not in mutants. The rate of aging decreased with osmotic pressure, supporting the idea that water is important for stabilizing the transition state. Molecular dynamics simulations were performed on the wtDIP adduct to relate the kinetic data to hydration changes in the enzyme active site gorge. The pH dependence of the melting temperature (Tm) of native and soman-wtBuChE, as determined by differential scanning calorimetry (DSC), indicated that the stabilization energy of aged BuChE is mainly due to the salt bridge between protonated H438 and PO-, with pK(H438) = 8.3. Electrophoresis under high pressure up to 2.5 kbar showed that aged wtBuChE did not undergo pressure-induced molten globule transition unlike the native enzyme. This transition was not seen for the mutant enzymes, indicating that mutants are resistant to the penetration of water into their structure. Our results support the conclusion that D70 and E197 are major residues for the water/H-bond network dynamics in the active site gorge of BuChE, both residues acting like valves. In mutant enzymes, mutated residues function like check valves: forced penetration of water in the gorge is difficult, release of water is facilitated. Study holds ProTherm entries: 12944, 12945 Extra Details: butyrylcholinesterase; organophosphate inhibitor; aging; hydorophobic pressure;,osmotic pressure; molecular dynamics

Submission Details

ID: JsBfRcXq

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:44 p.m.

Version: 1

Publication Details
Masson P;Fortier PL;Albaret C;Cléry C;Guerra P;Lockridge O,Chem. Biol. Interact. (1999) Structural and hydration changes in the active site gorge of phosporhylated butyrylcholinesterase accompanying the aging process. PMID:10421435
Additional Information

Study Summary

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
6QAA 2019-03-27 1.9 Human Butyrylcholinesterase in complex with (S)-2-(butylamino)-N-(2-cycloheptylethyl)-3-(1H-indol-3-yl)propanamide
2WSL 2009-09-15 2.0 Aged Form of Human Butyrylcholinesterase Inhibited by Tabun Analogue TA4
1P0I 2003-08-05 2.0 Crystal structure of human butyryl cholinesterase
2XQF 2011-03-23 2.1 X-ray Structure of human butyrylcholinesterase inhibited by racemic VX
2WIK 2009-05-19 2.1 NONAGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA6
5NN0 2018-03-14 2.1 Crystal structure of huBChE with N-((1-(2,3-dihydro-1H-inden-2-yl)piperidin-3-yl)methyl)-N-(2-(dimethylamino)ethyl)-2-naphthamide.
2WIJ 2009-05-19 2.1 NONAGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA5
2XMB 2010-12-01 2.1 G117H mutant of human butyrylcholinesterase in complex with sulfate
4BDS 2013-05-29 2.1 Human butyrylcholinesterase in complex with tacrine
3DJY 2008-12-02 2.1 Nonaged Form of Human Butyrylcholinesterase Inhibited by Tabun
1XLW 2005-02-01 2.1 Diethylphosphorylated Butyrylcholinesterase (Nonaged) Obtained By Reaction With Echothiophate
2WIG 2009-05-19 2.15 NONAGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA4
1XLU 2005-02-01 2.2 X-Ray Structure Of Di-Isopropyl-Phosphoro-Fluoridate (Dfp) Inhibited Butyrylcholinesterase after Aging
2WIF 2009-05-19 2.25 AGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA1
1XLV 2005-02-01 2.25 Ethylphosphorylated Butyrylcholinesterase (Aged) Obtained By Reaction With Echothiophate
6EP4 2017-12-13 2.3 Human butyrylcholinesterase in complex with decamethonium
2WID 2009-05-19 2.3 NONAGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA1
2XQG 2011-03-23 2.3 X-ray Structure of human butyrylcholinesterase inhibited by racemic VR
1P0P 2003-08-05 2.3 Crystal structure of soman-aged human butyryl cholinesterase in complex with the substrate analog butyrylthiocholine
2XMD 2010-12-01 2.3 G117H mutant of human butyrylcholinesterase in complex with echothiophate
3DKK 2008-12-02 2.31 Aged Form of Human Butyrylcholinesterase Inhibited by Tabun
6EQP 2017-12-13 2.35 Human butyrylcholinesterase in complex with ethopropazine
4B0O 2012-08-29 2.35 Crystal structure of soman-aged human butyrylcholinesterase in complex with benzyl pyridinium-4-methyltrichloroacetimidate
6I0B 2019-03-27 2.38 Human butyrylcholinesterase in complex with the S enantiomer of a chlorotacrine-tryptophan multi-target inhibitor.
1P0M 2003-08-05 2.38 Crystal structure of human butyryl cholinesterase in complex with a choline molecule
6EQQ 2017-12-13 2.4 Human butyrylcholinesterase in complex with huprine 19
2XMC 2010-12-01 2.4 G117H mutant of human butyrylcholinesterase in complex with fluoride anion
2XQK 2011-03-23 2.4 X-ray Structure of human butyrylcholinesterase inhibited by pure enantiomer VX-(S)
2XQJ 2011-03-23 2.4 X-ray Structure of human butyrylcholinesterase inhibited by pure enantiomer VX-(R)
4AXB 2012-08-29 2.4 Crystal structure of soman-aged human butyrylcholinesterase in complex with 2-PAM
1P0Q 2003-08-05 2.43 Crystal structure of soman-aged human butyryl cholinesterase
6EMI 2017-11-08 2.48 Crystal structure of a variant of human butyrylcholinesterase expressed in bacteria.
6QAB 2019-03-27 2.49 Human Butyrylcholinesterase in complex with (S)-N-(1-((2-cycloheptylethyl)amino)-3-(1H-indol-3-yl)-1-oxopropan-2-yl)-N,N-dimethylbutan-1-aminium
6QAE 2019-03-27 2.49 Human Butyrylcholinesterase in complex with (S)-N2-butyl-N1-(2-cycloheptylethyl)-3-(1H-indol-3-yl)-N1,N2-dimethylpropane-1,2-diamine
4AQD 2012-07-04 2.5 Crystal structure of fully glycosylated human butyrylcholinesterase
2Y1K 2011-06-29 2.5 STRUCTURE OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY CBDP (12H SOAK): PHOSPHOSERINE ADDUCT
6QAD 2019-03-27 2.5 Human Butyrylcholinesterase in complex with ((S)-2-(butylamino)-N-(2-(4-(dimethylamino)cyclohexyl)ethyl)-3-(1H-indol-3-yl)propanamide
4B0P 2012-08-29 2.5 Crystal structure of soman-aged human butyrylcholinesterase in complex with methyl 2-(pentafluorobenzyloxyimino)pyridinium
5DYW 2017-01-18 2.5 Crystal structure of human butyrylcholinesterase in complex with N-((1-benzylpiperidin-3-yl)methyl)-N-(2-methoxyethyl)naphthalene-2-sulfonamide
5LKR 2016-12-14 2.52 Human Butyrylcholinesterase complexed with N-Propargyliperidines
5DYT 2016-10-05 2.55 Crystal structure of human butyrylcholinesterase in complex with N-((1-benzylpiperidin-3-yl)methyl)-N-methylnaphthalene-2-sulfonamide
2XQI 2011-03-23 2.6 X-ray Structure of human butyrylcholinesterase inhibited by racemic CVX
6EYF 2018-11-21 2.6 Butyrylcolinesterase expressed in CHO cells co-crystallised with a rivastigmine analogue
6EUL 2018-11-14 2.6 Butyrylcolinesterase expressed in CHO cells co-crystallised with a rivastigmine analogue
6F7Q 2018-09-05 2.6 Human Butyrylcholinesterase complexed with N-Propargyliperidines
5DYY 2017-01-18 2.65 Crystal structure of human butyrylcholinesterase in complex with N-((1-benzylpiperidin-3-yl)methyl)naphthalene-2-sulfonamide
6I0C 2019-03-27 2.67 Human butyrylcholinesterase in complex with the R enantiomer of a chlorotacrine-tryptophan multi-target inhibitor.
6EZ2 2018-12-12 2.7 Human butyrylcholinesterase carbamylated.
4XII 2015-07-08 2.7 X-ray structure of human butyrylcholinesterase in complex with N-((1-(2,3-dihydro-1H-inden-2-yl)piperidin-3-yl)methyl)-8-hydroxy-N-(2-methoxyethyl)-5-nitroquinoline-7-carboxamide
4BBZ 2013-02-06 2.7 Structure of human butyrylcholinesterase inhibited by CBDP (2-min soak): Cresyl-phosphoserine adduct
4TPK 2014-10-22 2.7 Human butyrylcholinesterase in complex with N-((1-(2,3-dihydro-1H-inden-2-yl)piperidin-3-yl)methyl)-N-(2-methoxyethyl)-2-naphthamide
2XMG 2010-12-01 2.7 G117H mutant of human butyrylcholinesterase in complex with VX
2J4C 2007-03-27 2.75 Structure of human Butyrylcholinesterase in complex with 10mM HgCl2
6QAC 2019-03-27 2.77 Human Butyrylcholinesterase in complex with (S)-2-(butylamino)-N-(3-cycloheptylpropyl)-3-(1H-indol-3-yl)propanamide
5K5E 2016-07-27 2.8 Discovery and Structure-Activity Relationships of a Highly Selective Butyrylcholinesterase Inhibitor by Structure-Based Virtual Screening
6ESY 2017-12-13 2.8 Human butyrylcholinesterase in complex with thioflavine T
2PM8 2007-09-25 2.8 Crystal structure of recombinant full length human butyrylcholinesterase
6ESJ 2017-12-13 2.98 Human butyrylcholinesterase in complex with propidium
3O9M 2011-04-13 2.98 Co-crystallization studies of full length recombinant BChE with cocaine offers insights into cocaine detoxification
2WIL 2009-05-26 3.1 AGED FORM OF HUMAN BUTYRYLCHOLINESTERASE INHIBITED BY TABUN ANALOGUE TA5
6I2T 2018-12-19 5.7 CryoEM reconstruction of full-length, fully-glycosylated human butyrylcholinesterase tetramer

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.3 BCHE protein P21927 CHLE_RABIT
95.0 BCHE protein P81908 CHLE_HORSE
92.1 BCHE protein O62761 CHLE_PANTT
92.1 BCHE protein O62760 CHLE_FELCA
92.1 BCHE protein P32749 CHLE_BOVIN
100.0 BCHE protein P06276 CHLE_HUMAN