4hpa - Revision history

OCA at 14:09, 8 November 2023

OCA — Wed, 08 Nov 2023 14:09:31 GMT

←Older revision		Revision as of 14:09, 8 November 2023
Line 4:		Line 4:
	== Structural highlights ==		== Structural highlights ==
	<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodnius_prolixus Rhodnius prolixus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4HPA FirstGlance]. <br>		<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodnius_prolixus Rhodnius prolixus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4HPA FirstGlance]. <br>
-	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</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]] 1.5Å</td></tr>
		+	<tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [https://pdbe.org/4hpa PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [https://www.ebi.ac.uk/pdbsum/4hpa PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4hpa ProSAT]</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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [https://pdbe.org/4hpa PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [https://www.ebi.ac.uk/pdbsum/4hpa PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4hpa ProSAT]</span></td></tr>
	</table>		</table>

OCA at 08:30, 3 November 2022

OCA — Thu, 03 Nov 2022 08:30:51 GMT

←Older revision		Revision as of 08:30, 3 November 2022
Line 3:		Line 3:
	<StructureSection load='4hpa' size='340' side='right'caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>		<StructureSection load='4hpa' size='340' side='right'caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
-	<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [~~http~~://en.wikipedia.org/wiki/~~Rhopr Rhopr~~]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [~~http~~://~~oca~~.~~weizmann.ac.il/oca-docs~~/fgij/fg.htm?mol=4HPA FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodnius_prolixus Rhodnius prolixus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4HPA FirstGlance]. <br>
-	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene>~~</td></tr>~~	+	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>
-	~~<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[4hpb\|4hpb]], [[4hpc\|4hpc]], [[4hpd\|4hpd]]~~</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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [https://pdbe.org/4hpa PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [https://www.ebi.ac.uk/pdbsum/4hpa PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4hpa 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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [~~http~~://pdbe.org/4hpa PDBe], [~~http~~://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [~~http~~://www.ebi.ac.uk/pdbsum/4hpa PDBsum], [~~http~~://prosat.h-its.org/prosat/prosatexe?pdbcode=4hpa ProSAT]</span></td></tr>	+
	</table>		</table>
	== Function ==		== Function ==
-	[~~[http~~://www.uniprot.org/uniprot/NP4_RHOPR NP4_RHOPR]] Heme-based protein that deliver nitric oxide gas (NO) to the victim while feeding, resulting in vasodilation and inhibition of platelet aggregation. Also bind tightly to histamine, which is released by the host to induce wound healing (By similarity).	+	[https://www.uniprot.org/uniprot/NP4_RHOPR NP4_RHOPR] Heme-based protein that deliver nitric oxide gas (NO) to the victim while feeding, resulting in vasodilation and inhibition of platelet aggregation. Also bind tightly to histamine, which is released by the host to induce wound healing (By similarity).
	<div style="background-color:#fffaf0;">		<div style="background-color:#fffaf0;">
	== Publication Abstract from PubMed ==		== Publication Abstract from PubMed ==
Line 27:		Line 26:
	</StructureSection>		</StructureSection>
	[[Category: Large Structures]]		[[Category: Large Structures]]
-	[[Category: ~~Rhopr~~]]	+	[[Category: Rhodnius prolixus]]
-	[[Category: Knipp, M]]	+	[[Category: Knipp M]]
-	[[Category: Nishikawa, K]]	+	[[Category: Nishikawa K]]
-	[[Category: Ogata, H]]	+	[[Category: Ogata H]]
-	~~[[Category: Heme]]~~	+
-	~~[[Category: Lipocalin]]~~	+
-	~~[[Category: Metal binding protein]]~~	+
-	~~[[Category: Nitrophorin]]~~	+
-	~~[[Category: Sulfide]]~~	+
-	~~[[Category: Transport protein~~]]	+

OCA at 08:10, 27 March 2019

OCA — Wed, 27 Mar 2019 08:10:15 GMT

←Older revision		Revision as of 08:10, 27 March 2019
Line 1:		Line 1:

	==Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4==		==Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4==
-	<StructureSection load='4hpa' size='340' side='right' caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>	+	<StructureSection load='4hpa' size='340' side='right'caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
	<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhopr Rhopr]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4HPA FirstGlance]. <br>		<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhopr Rhopr]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4HPA FirstGlance]. <br>
Line 19:		Line 19:
	</div>		</div>
	<div class="pdbe-citations 4hpa" style="background-color:#fffaf0;"></div>		<div class="pdbe-citations 4hpa" style="background-color:#fffaf0;"></div>
		+
		+	==See Also==
		+	*[[Nitrophorin\|Nitrophorin]]
	== References ==		== References ==
	<references/>		<references/>
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
		+	[[Category: Large Structures]]
	[[Category: Rhopr]]		[[Category: Rhopr]]
	[[Category: Knipp, M]]		[[Category: Knipp, M]]

OCA at 21:57, 5 August 2016

OCA — Fri, 05 Aug 2016 21:57:24 GMT

←Older revision		Revision as of 21:57, 5 August 2016
Line 1:		Line 1:
		+
	==Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4==		==Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4==
	<StructureSection load='4hpa' size='340' side='right' caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>		<StructureSection load='4hpa' size='340' side='right' caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>
Line 5:		Line 6:
	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>		</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>
	<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[4hpb\|4hpb]], [[4hpc\|4hpc]], [[4hpd\|4hpd]]</td></tr>		<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[4hpb\|4hpb]], [[4hpc\|4hpc]], [[4hpd\|4hpd]]</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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [http://www.ebi.ac.uk/pdbsum/4hpa PDBsum]</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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [http://pdbe.org/4hpa PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [http://www.ebi.ac.uk/pdbsum/4hpa PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4hpa ProSAT]</span></td></tr>
	</table>		</table>
	== Function ==		== Function ==
Line 17:		Line 18:
	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>		From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
	</div>		</div>
		+	<div class="pdbe-citations 4hpa" style="background-color:#fffaf0;"></div>
	== References ==		== References ==
	<references/>		<references/>

OCA at 06:10, 25 December 2014

OCA — Thu, 25 Dec 2014 06:10:35 GMT

←Older revision		Revision as of 06:10, 25 December 2014
Line 1:		Line 1:
-	~~{{STRUCTURE_4hpa\| PDB=4hpa \| SCENE= }}~~	+	==Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4==
-	===Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4===	+	<StructureSection load='4hpa' size='340' side='right' caption='[[4hpa]], [[Resolution\|resolution]] 1.50Å' scene=''>
-	~~{{ABSTRACT_PUBMED_23474537}}~~	+	== Structural highlights ==
-		+	<table><tr><td colspan='2'>[[4hpa]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhopr Rhopr]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4HPA FirstGlance]. <br>
-	==Function==	+	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=H2S:HYDROSULFURIC+ACID'>H2S</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene></td></tr>
		+	<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[4hpb\|4hpb]], [[4hpc\|4hpc]], [[4hpd\|4hpd]]</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=4hpa FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4hpa OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4hpa RCSB], [http://www.ebi.ac.uk/pdbsum/4hpa PDBsum]</span></td></tr>
		+	</table>
		+	== Function ==
	[[http://www.uniprot.org/uniprot/NP4_RHOPR NP4_RHOPR]] Heme-based protein that deliver nitric oxide gas (NO) to the victim while feeding, resulting in vasodilation and inhibition of platelet aggregation. Also bind tightly to histamine, which is released by the host to induce wound healing (By similarity).		[[http://www.uniprot.org/uniprot/NP4_RHOPR NP4_RHOPR]] Heme-based protein that deliver nitric oxide gas (NO) to the victim while feeding, resulting in vasodilation and inhibition of platelet aggregation. Also bind tightly to histamine, which is released by the host to induce wound healing (By similarity).
		+	<div style="background-color:#fffaf0;">
		+	== Publication Abstract from PubMed ==
		+	Nitrophorins are proteins occurring in the saliva of the blood-sucking insect Rhodnius prolixus to carry NO as a vasodilator and blood-coagulation inhibitor into the victim's tissue. It was suggested that the rate of NO release can be enhanced by the blood-plasma component L-cysteine [J.M.C.Ribeiro, Insect Biochem. Mol. Biol. 26 (1996) 899-905]. However, the mechanism of the reaction is not clear. In the attempt to exploit the reaction in detail, complexes of nitrophorin 4 (NP4) with the thiols 2-mercaptoethanol, L-cysteine, and L-homocysteine and with HS(-) were formed and characterized under anaerobic conditions using absorption spectroscopy, X-ray crystallography, and EPR spectroscopy. In contrast to met-myoglobin, which is reduced by L-cysteine, all four compounds form low-spin Fe(III) complexes with NP4. The weak equilibration constants (167-5200 M(-1)) neither support significant complexation nor the simple displacement of NO in vivo. Both amino acid based thiols form additional H-bonds with side chains of the heme pocket entry. Glutathione and L-methionine did not form a complex, indicating the specificity of the complexes with L-cysteine and L-homocysteine. Continuous wave EPR spectroscopy reveals the simultaneous existence of three low-spin systems in each case that are attributed to various protonation and/or conformational stages in the heme pocket. Electron nuclear double resonance (ENDOR) spectroscopy demonstrates that the thiol sulfurs are, at least in part, protonated. Overall, the results not only demonstrate the good accessibility of the NP4 heme center by biologically relevant thiols, but also represent the first structural characterization of a ferriheme protein in complex with L-cysteine L-homocysteine.

-	~~==About this Structure==~~	+	Complexes of ferriheme nitrophorin 4 with low-molecular weight thiol(ate)s occurring in blood plasma.,He C, Nishikawa K, Erdem OF, Reijerse E, Ogata H, Lubitz W, Knipp M J Inorg Biochem. 2013 May;122:38-48. doi: 10.1016/j.jinorgbio.2013.01.012. Epub, 2013 Feb 4. PMID:23474537<ref>PMID:23474537</ref>
-	~~[[4hpa]] is a 1 chain structure~~ with ~~sequence from [http://en~~.~~wikipedia~~.~~org/wiki/Rhopr Rhopr]~~. ~~Full crystallographic information is available from [http~~:/~~/oca~~.~~weizmann~~.ac.~~il/oca-bin/ocashort?id=4HPA OCA]~~.	+

-	==~~Reference~~==	+	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
-	~~<ref group="xtra">PMID:023474537</ref>~~<references ~~group="xtra"~~/><~~references~~/>	+	</div>
		+	== References ==
		+	<references/>
		+	__TOC__
		+	</StructureSection>
	[[Category: Rhopr]]		[[Category: Rhopr]]
-	[[Category: Knipp, M.]]	+	[[Category: Knipp, M]]
-	[[Category: Nishikawa, K.]]	+	[[Category: Nishikawa, K]]
-	[[Category: Ogata, H.]]	+	[[Category: Ogata, H]]
	[[Category: Heme]]		[[Category: Heme]]
	[[Category: Lipocalin]]		[[Category: Lipocalin]]

OCA at 08:45, 19 March 2014

OCA — Wed, 19 Mar 2014 08:45:57 GMT

←Older revision		Revision as of 08:45, 19 March 2014
Line 1:		Line 1:
-	~~'''Unreleased~~ structure~~'''~~	+	{{STRUCTURE_4hpa\| PDB=4hpa \| SCENE= }}
		+	===Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4===
		+	{{ABSTRACT_PUBMED_23474537}}

-	~~The entry 4hpa~~ is ~~ON HOLD until Paper Publication~~	+	==Function==
		+	[[http://www.uniprot.org/uniprot/NP4_RHOPR NP4_RHOPR]] Heme-based protein that deliver nitric oxide gas (NO) to the victim while feeding, resulting in vasodilation and inhibition of platelet aggregation. Also bind tightly to histamine, which is released by the host to induce wound healing (By similarity).

-	~~Authors~~: ~~Nishikawa, K~~.~~, Ogata, H~~.~~, Knipp, M~~.	+	==About this Structure==
		+	[[4hpa]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhopr Rhopr]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4HPA OCA].

-	~~Description~~: ~~Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7~~.4	+	==Reference==
		+	<ref group="xtra">PMID:023474537</ref><references group="xtra"/><references/>
		+	[[Category: Rhopr]]
		+	[[Category: Knipp, M.]]
		+	[[Category: Nishikawa, K.]]
		+	[[Category: Ogata, H.]]
		+	[[Category: Heme]]
		+	[[Category: Lipocalin]]
		+	[[Category: Metal binding protein]]
		+	[[Category: Nitrophorin]]
		+	[[Category: Sulfide]]
		+	[[Category: Transport protein]]

OCA at 13:42, 6 March 2013

OCA — Wed, 06 Mar 2013 13:42:38 GMT

←Older revision		Revision as of 13:42, 6 March 2013
Line 1:		Line 1:
	'''Unreleased structure'''		'''Unreleased structure'''

-	The entry 4hpa is ON HOLD	+	The entry 4hpa is ON HOLD until Paper Publication

	Authors: Nishikawa, K., Ogata, H., Knipp, M.		Authors: Nishikawa, K., Ogata, H., Knipp, M.

	Description: Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4		Description: Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4

OCA: Protected "4hpa" [edit=sysop:move=sysop]

OCA — Wed, 31 Oct 2012 08:43:16 GMT

Protected "4hpa" [edit=sysop:move=sysop]

←Older revision

Revision as of 08:43, 31 October 2012

OCA: New page: '''Unreleased structure''' The entry 4hpa is ON HOLD Authors: Nishikawa, K., Ogata, H., Knipp, M. Description: Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with ...

OCA — Wed, 31 Oct 2012 08:43:15 GMT

New page: '''Unreleased structure''' The entry 4hpa is ON HOLD Authors: Nishikawa, K., Ogata, H., Knipp, M. Description: Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with ...

New page

'''Unreleased structure'''

The entry 4hpa is ON HOLD

Authors: Nishikawa, K., Ogata, H., Knipp, M.

Description: Crystal structure of Nitrophorin 4 from Rhodnius prolixus Complexed with sulfide ion at pH 7.4