5toy - Revision history

OCA at 13:07, 4 October 2023

OCA — Wed, 04 Oct 2023 13:07:01 GMT

←Older revision		Revision as of 13:07, 4 October 2023
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	<StructureSection load='5toy' size='340' side='right'caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>		<StructureSection load='5toy' size='340' side='right'caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
-	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure with sequence from [~~http~~://en.wikipedia.org/wiki/~~"bacillus_coli"_migula_1895 "bacillus~~ coli~~" migula 1895~~]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [~~http~~://~~oca~~.~~weizmann.ac.il/oca-docs~~/fgij/fg.htm?mol=5TOY FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5TOY FirstGlance]. <br>
-	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=JSC:[(1,2,3,4,5-ETA)-1-(4-{[CARBOXY(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSC</scene>, <scene name='pdbligand=JSD:[(1,2,3,4,5-ETA)-1-(4-{[(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSD</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=RU:~~RUTHENIUM+ION~~'>RU</scene>, <scene name='pdbligand=~~SUC~~:~~SUCROSE~~'>~~SUC~~</scene~~></td></tr>~~	+	</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models\|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution\|Resolution]] 1.3Å</td></tr>
-	<tr id='gene'><td class="sblockLbl"><b>[[Gene\|Gene:]]</b></td><td class="sblockDat">blaCTX-M, CTX-M-14 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=562 "Bacillus coli" Migula 1895])</td></tr>	+	<tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FRU:FRUCTOSE'>FRU</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=JSC:[(1,2,3,4,5-ETA)-1-(4-{[CARBOXY(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSC</scene>, <scene name='pdbligand=JSD:[(1,2,3,4,5-ETA)-1-(4-{[(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSD</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=PRD_900003:sucrose'>PRD_900003</scene>, <scene name='pdbligand=RU:RUTHENIUM+ION'>RU</scene></td></tr>
-	<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-lactamase Beta-lactamase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.6 3.5.2.6] </span></td></tr>	+	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5toy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5toy OCA], [https://pdbe.org/5toy PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5toy RCSB], [https://www.ebi.ac.uk/pdbsum/5toy PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5toy ProSAT]</span></td></tr>
-	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[~~http~~://~~oca~~.~~weizmann.ac.il/oca-docs~~/fgij/fg.htm?mol=5toy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5toy OCA], [~~http~~://pdbe.org/5toy PDBe], [~~http~~://www.rcsb.org/pdb/explore.do?structureId=5toy RCSB], [~~http~~://www.ebi.ac.uk/pdbsum/5toy PDBsum], [~~http~~://prosat.h-its.org/prosat/prosatexe?pdbcode=5toy ProSAT]</span></td></tr>	+
	</table>		</table>
		+	== Function ==
		+	[https://www.uniprot.org/uniprot/Q9L5C7_ECOLX Q9L5C7_ECOLX]
	<div style="background-color:#fffaf0;">		<div style="background-color:#fffaf0;">
	== Publication Abstract from PubMed ==		== Publication Abstract from PubMed ==
Line 25:		Line 26:
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
-	[[Category: ~~Bacillus~~ coli ~~migula 1895]]~~	+	[[Category: Escherichia coli]]
-	~~[[Category: Beta-lactamase~~]]	+
	[[Category: Large Structures]]		[[Category: Large Structures]]
-	[[Category: Chen, Y]]	+	[[Category: Chen Y]]
-	[[Category: Lewandowski~~, E M]]~~	+	[[Category: Lewandowski EM]]
-	~~[[Category: Ctx-m-14]]~~	+
-	~~[[Category: Hydrolase-antibiotic complex]]~~	+
-	~~[[Category: Organometallic]]~~	+
-	~~[[Category: Ruthenocene~~]]	+

OCA at 16:57, 11 December 2019

OCA — Wed, 11 Dec 2019 16:57:38 GMT

←Older revision		Revision as of 16:57, 11 December 2019
Line 1:		Line 1:

	==X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Products in Complex with CTX-M-14 E166A Beta-Lactamase==		==X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Products in Complex with CTX-M-14 E166A Beta-Lactamase==
-	<StructureSection load='5toy' size='340' side='right' caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>	+	<StructureSection load='5toy' size='340' side='right'caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacillus_coli"_migula_1895 "bacillus coli" migula 1895]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5TOY FirstGlance]. <br>		<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacillus_coli"_migula_1895 "bacillus coli" migula 1895]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5TOY FirstGlance]. <br>
Line 18:		Line 18:
	</div>		</div>
	<div class="pdbe-citations 5toy" style="background-color:#fffaf0;"></div>		<div class="pdbe-citations 5toy" style="background-color:#fffaf0;"></div>
		+
		+	==See Also==
		+	*[[Beta-lactamase 3D structures\|Beta-lactamase 3D structures]]
	== References ==		== References ==
	<references/>		<references/>
Line 24:		Line 27:
	[[Category: Bacillus coli migula 1895]]		[[Category: Bacillus coli migula 1895]]
	[[Category: Beta-lactamase]]		[[Category: Beta-lactamase]]
		+	[[Category: Large Structures]]
	[[Category: Chen, Y]]		[[Category: Chen, Y]]
	[[Category: Lewandowski, E M]]		[[Category: Lewandowski, E M]]

OCA at 10:14, 15 November 2017

OCA — Wed, 15 Nov 2017 10:14:29 GMT

←Older revision		Revision as of 10:14, 15 November 2017
Line 3:		Line 3:
	<StructureSection load='5toy' size='340' side='right' caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>		<StructureSection load='5toy' size='340' side='right' caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
-	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5TOY FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"bacillus_coli"_migula_1895 "bacillus coli" migula 1895]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5TOY FirstGlance]. <br>
	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=JSC:[(1,2,3,4,5-ETA)-1-(4-{[CARBOXY(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSC</scene>, <scene name='pdbligand=JSD:[(1,2,3,4,5-ETA)-1-(4-{[(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSD</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=RU:RUTHENIUM+ION'>RU</scene>, <scene name='pdbligand=SUC:SUCROSE'>SUC</scene></td></tr>		</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=JSC:[(1,2,3,4,5-ETA)-1-(4-{[CARBOXY(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSC</scene>, <scene name='pdbligand=JSD:[(1,2,3,4,5-ETA)-1-(4-{[(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSD</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=RU:RUTHENIUM+ION'>RU</scene>, <scene name='pdbligand=SUC:SUCROSE'>SUC</scene></td></tr>
		+	<tr id='gene'><td class="sblockLbl"><b>[[Gene\|Gene:]]</b></td><td class="sblockDat">blaCTX-M, CTX-M-14 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=562 "Bacillus coli" Migula 1895])</td></tr>
	<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-lactamase Beta-lactamase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.6 3.5.2.6] </span></td></tr>		<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-lactamase Beta-lactamase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.6 3.5.2.6] </span></td></tr>
	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5toy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5toy OCA], [http://pdbe.org/5toy PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5toy RCSB], [http://www.ebi.ac.uk/pdbsum/5toy PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5toy ProSAT]</span></td></tr>		<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5toy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5toy OCA], [http://pdbe.org/5toy PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5toy RCSB], [http://www.ebi.ac.uk/pdbsum/5toy PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5toy ProSAT]</span></td></tr>
	</table>		</table>
		+	<div style="background-color:#fffaf0;">
		+	== Publication Abstract from PubMed ==
		+	The beta-lactam antibiotics inhibit penicillin-binding proteins (PBPs) by forming a stable, covalent, acyl-enzyme complex. During the evolution from PBPs to Class A beta-lactamases, the beta-lactamases acquired Glu166 to activate a catalytic water and cleave the acyl-enzyme bond. Here we present three product complex crystal structures of CTX-M-14 Class A beta-lactamase with a ruthenocene-conjugated penicillin - a 0.85 A resolution structure of E166A mutant complexed with the penilloate product, a 1.30 A resolution complex structure of the same mutant with the penicilloate product, and a 1.18 A resolution complex structure of S70G mutant with a penicilloate product epimer - shedding light on the catalytic mechanisms and product inhibition of PBPs and Class A beta-lactamases. The E166A-penilloate complex captured the hydrogen bonding network following the protonation of the leaving group and, for the first time, unambiguously show that the ring nitrogen donates a proton to Ser130, which in turn donates a proton to Lys73. These observations indicate that in the absence of Glu166, the equivalent lysine would be neutral in PBPs and therefore capable of serving as the general base to activate the catalytic serine. Together with previous results, this structure suggests a common proton relay network shared by Class A beta-lactamases and PBPs, from the catalytic serine to the lysine, and ultimately to the ring nitrogen. Additionally, the E166A-penicilloate complex reveals previously unseen conformational changes of key catalytic residues during the release of the product, and is the first structure to capture the hydrolyzed product in the presence of an unmutated catalytic serine. This article is protected by copyright. All rights reserved.
		+
		+	Mechanisms of Proton Relay and Product Release by Class A beta-Lactamase at Ultrahigh Resolution.,Lewandowski EM, Lethbridge KG, Sanishvili R, Skiba J, Kowalski K, Chen Y FEBS J. 2017 Nov 2. doi: 10.1111/febs.14315. PMID:29095570<ref>PMID:29095570</ref>
		+
		+	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
		+	</div>
		+	<div class="pdbe-citations 5toy" style="background-color:#fffaf0;"></div>
		+	== References ==
		+	<references/>
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
		+	[[Category: Bacillus coli migula 1895]]
	[[Category: Beta-lactamase]]		[[Category: Beta-lactamase]]
	[[Category: Chen, Y]]		[[Category: Chen, Y]]

OCA at 07:49, 8 November 2017

OCA — Wed, 08 Nov 2017 07:49:26 GMT

←Older revision		Revision as of 07:49, 8 November 2017
Line 1:		Line 1:
-	'''Unreleased structure'''

-	~~The entry 5toy is ON HOLD until Apr 18 2019~~	+	==X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Products in Complex with CTX-M-14 E166A Beta-Lactamase==
-		+	<StructureSection load='5toy' size='340' side='right' caption='[[5toy]], [[Resolution\|resolution]] 1.30Å' scene=''>
-	~~Authors: Lewandowski, E.M., Chen, Y.~~	+	== Structural highlights ==
-		+	<table><tr><td colspan='2'>[[5toy]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5TOY OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5TOY FirstGlance]. <br>
-	~~Description:~~ X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Products in Complex with CTX-M-14 E166A Beta-Lactamase	+	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=JSC:[(1,2,3,4,5-ETA)-1-(4-{[CARBOXY(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSC</scene>, <scene name='pdbligand=JSD:[(1,2,3,4,5-ETA)-1-(4-{[(4-CARBOXY-5,5-DIMETHYL-1,3-THIAZOLIDIN-2-YL)METHYL]AMINO}-4-OXOBUTANOYL)CYCLOPENTADIENYL][(1,2,3,4,5-ETA)-CYCLOPENTADIENYL]RUTHENIUM'>JSD</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene>, <scene name='pdbligand=RU:RUTHENIUM+ION'>RU</scene>, <scene name='pdbligand=SUC:SUCROSE'>SUC</scene></td></tr>
-	[[Category: ~~Unreleased Structures~~]]	+	<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Beta-lactamase Beta-lactamase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.2.6 3.5.2.6] </span></td></tr>
		+	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5toy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5toy OCA], [http://pdbe.org/5toy PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5toy RCSB], [http://www.ebi.ac.uk/pdbsum/5toy PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5toy ProSAT]</span></td></tr>
		+	</table>
		+	__TOC__
		+	</StructureSection>
		+	[[Category: Beta-lactamase]]
	[[Category: Chen, Y]]		[[Category: Chen, Y]]
-	[[Category: Lewandowski, E.M]]	+	[[Category: Lewandowski, E M]]
		+	[[Category: Ctx-m-14]]
		+	[[Category: Hydrolase-antibiotic complex]]
		+	[[Category: Organometallic]]
		+	[[Category: Ruthenocene]]

OCA at 13:26, 27 September 2017

OCA — Wed, 27 Sep 2017 13:26:07 GMT

←Older revision		Revision as of 13:26, 27 September 2017
Line 1:		Line 1:
	'''Unreleased structure'''		'''Unreleased structure'''

-	The entry 5toy is ON HOLD	+	The entry 5toy is ON HOLD until Apr 18 2019

	Authors: Lewandowski, E.M., Chen, Y.		Authors: Lewandowski, E.M., Chen, Y.

OCA: Protected "5toy" [edit=sysop:move=sysop]

OCA — Wed, 26 Oct 2016 17:20:00 GMT

Protected "5toy" [edit=sysop:move=sysop]

←Older revision

Revision as of 17:20, 26 October 2016

OCA: New page: '''Unreleased structure''' The entry 5toy is ON HOLD Authors: Lewandowski, E.M., Chen, Y. Description: X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Pro...

OCA — Wed, 26 Oct 2016 17:19:59 GMT

New page: '''Unreleased structure''' The entry 5toy is ON HOLD Authors: Lewandowski, E.M., Chen, Y. Description: X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Pro...

New page

'''Unreleased structure'''

The entry 5toy is ON HOLD

Authors: Lewandowski, E.M., Chen, Y.

Description: X-Ray Crystal Structure of Ruthenocene Conjugated Penicilloate and Penilloate Products in Complex with CTX-M-14 E166A Beta-Lactamase
[[Category: Unreleased Structures]]
[[Category: Chen, Y]]
[[Category: Lewandowski, E.M]]