2pkj - Revision history

OCA: Redirecting to 3mh0

OCA — Wed, 21 Apr 2010 06:51:56 GMT

Redirecting to 3mh0

←Older revision		Revision as of 06:51, 21 April 2010
Line 1:		Line 1:
-	~~{{Seed}}~~	+	#REDIRECT [[3mh0]] This PDB entry is obsolete and replaced by 3mh0
-	[[~~Image:2pkj.png\|left\|200px~~]]	+
-		+
-	~~<!--~~	+
-	~~The line below this paragraph, containing "STRUCTURE_2pkj", creates the "Structure Box" on the page.~~	+
-	~~You may change the~~ PDB ~~parameter (which sets the PDB file loaded into the applet)~~	+
-	~~or the SCENE parameter (which sets the initial scene displayed when the page~~ is ~~loaded),~~	+
-	~~or leave the SCENE parameter empty for the default display.~~	+
-	~~-->~~	+
-	~~{{STRUCTURE_2pkj\| PDB=2pkj \| SCENE= }}~~	+
-		+
-	~~===Crystal Structure of F169A p38 Kinase===~~	+
-		+
-		+
-	~~<!--~~	+
-	~~The line below this paragraph, {{ABSTRACT_PUBMED_17441692}}, adds the Publication Abstract to the page~~	+
-	~~(as it appears on PubMed at http://www.pubmed.gov), where 17441692 is the PubMed ID number.~~	+
-	~~-->~~	+
-	~~{{ABSTRACT_PUBMED_17441692}}~~	+
-		+
-	~~==About this Structure==~~	+
-	2PKJ is a 1 chain structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].	+
-		+
-	~~==Reference==~~	+
-	~~<ref group="xtra">PMID:17441692</ref><references group="xtra"/>~~	+
-	~~[[Category: Homo sapiens]]~~	+
-	~~[[Category: Mitogen-activated protein kinase]]~~	+
-	~~[[Category: Bukhtiyarova, M.]]~~	+
-	~~[[Category: Karpusas, M.]]~~	+
-	~~[[Category: Namboodiri, H V.M.]]~~	+
-	~~[[Category: Springman, E B.]]~~	+
-	~~[[Category: Protein kinase]]~~	+
-		+
-	~~''Page seeded~~ by ~~[http://oca.weizmann.ac.il/oca OCA ] on Tue Feb 17 14:25:02 2009''~~	+

OCA at 12:25, 17 February 2009

OCA — Tue, 17 Feb 2009 12:25:04 GMT

←Older revision		Revision as of 12:25, 17 February 2009
Line 20:		Line 20:

	==About this Structure==		==About this Structure==
-	2PKJ is a ~~[[Single protein]]~~ structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].	+	2PKJ is a 1 chain structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].

	==Reference==		==Reference==
-	Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state., Bukhtiyarova M, Karpusas M, Northrop K, Namboodiri HV, Springman EB, Biochemistry. 2007 May 15;46(19):5687-96. Epub 2007 Apr 19. PMID:~~[http:~~//~~www.ncbi.nlm.nih.gov/pubmed/17441692 17441692]~~	+	<ref group="xtra">PMID:17441692</ref><references group="xtra"/>
	[[Category: Homo sapiens]]		[[Category: Homo sapiens]]
	[[Category: Mitogen-activated protein kinase]]		[[Category: Mitogen-activated protein kinase]]
-	[[Category: Single protein]]
	[[Category: Bukhtiyarova, M.]]		[[Category: Bukhtiyarova, M.]]
	[[Category: Karpusas, M.]]		[[Category: Karpusas, M.]]
Line 33:		Line 32:
	[[Category: Protein kinase]]		[[Category: Protein kinase]]

-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on ~~Mon Jul 28 19~~:49:~~16 2008~~''	+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Feb 17 14:25:02 2009''

OCA at 16:49, 28 July 2008

OCA — Mon, 28 Jul 2008 16:49:20 GMT

←Older revision		Revision as of 16:49, 28 July 2008
Line 1:		Line 1:
-	[[Image:2pkj.~~jpg~~\|left\|200px]]	+	{{Seed}}
		+	[[Image:2pkj.png\|left\|200px]]

	<!--		<!--
Line 9:		Line 10:
	{{STRUCTURE_2pkj\| PDB=2pkj \| SCENE= }}		{{STRUCTURE_2pkj\| PDB=2pkj \| SCENE= }}

-	~~'''~~Crystal Structure of F169A p38 Kinase~~'''~~	+	===Crystal Structure of F169A p38 Kinase===


-	~~==Overview==~~	+	<!--
-	~~In order to study the role of Phe169 in p38alpha MAP kinase structure and function~~, ~~wild-type p38alpha and five p38alpha DFG motif mutants were examined in vitro for phosphorylation by MKK6~~, kinase activity toward ATF2 substrate, thermal stability, and X-ray crystal structure. All six p38alpha variants were efficiently phosphorylated by MKK6. However, only one activated p38alpha mutant (F169Y) possessed measurable kinase activity (1% compared to wild-type). The loss of kinase activity among the ~~DFG mutants may result from an inability~~ to ~~correctly position Asp168 in~~ the ~~activated form of p38alpha. Two mutations significantly increased the thermal stability of p38alpha~~ (~~F169A DeltaTm = 1~~.~~3 degrees C and D168G DeltaTm = 3~~.~~8 degrees C~~), and two mutations significantly decreased the stability of p38alpha (F169R DeltaTm = -3.2 degrees C and F169G DeltaTm = -4.7 degrees C). Interestingly, X-ray crystal structures of two thermally destabilized p38alpha-F169R and p38alpha-F169G mutants revealed a DFG-OUT conformation in the absence of an inhibitor molecule. This DFG-OUT conformation, termed alpha-DFG-OUT, is ~~different from~~ the ~~ones previously identified in p38alpha crystal structures with bound inhibitors and postulated from high-temperature molecular dynamics simulations~~. ~~Taken together, these results indicate that Phe169 is optimized for p38alpha functional activity and structural dynamics, rather than for structural stability. The alpha~~-~~DFG~~-OUT conformation observed for p38alpha-F169R and p38alpha-F169G may represent a naturally occurring intermediate state of p38alpha that provides access for binding of allosteric inhibitors. A model of the local forces driving the DFG IN-OUT transition in p38alpha is proposed.	+	The line below this paragraph, {{ABSTRACT_PUBMED_17441692}}, adds the Publication Abstract to the page
		+	(as it appears on PubMed at http://www.pubmed.gov), where 17441692 is the PubMed ID number.
		+	-->
		+	{{ABSTRACT_PUBMED_17441692}}

	==About this Structure==		==About this Structure==
Line 28:		Line 32:
	[[Category: Springman, E B.]]		[[Category: Springman, E B.]]
	[[Category: Protein kinase]]		[[Category: Protein kinase]]
-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on ~~Sun May 4 13~~:18:19 2008''	+
		+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 19:49:16 2008''

OCA at 10:18, 4 May 2008

OCA — Sun, 04 May 2008 10:18:21 GMT

←Older revision		Revision as of 10:18, 4 May 2008
Line 1:		Line 1:
	[[Image:2pkj.jpg\|left\|200px]]		[[Image:2pkj.jpg\|left\|200px]]

-	~~{{Structure~~	+	<!--
-	~~\|PDB= 2pkj \|SIZE=350\|CAPTION= <scene name='initialview01'>2pkj</scene>~~, ~~resolution 2~~.~~000Å~~	+	The line below this paragraph, containing "STRUCTURE_2pkj", creates the "Structure Box" on the page.
-	~~\|SITE=~~	+	You may change the PDB parameter (which sets the PDB file loaded into the applet)
-	~~\|LIGAND= <~~scene ~~name='pdbligand=BOG:B-OCTYLGLUCOSIDE'>BOG</scene>~~	+	or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
-	~~\|ACTIVITY= <span class='plainlinks'>[http://en~~.~~wikipedia.org/wiki/Mitogen~~-~~activated_protein_kinase Mitogen~~-~~activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] </span~~>	+	or leave the SCENE parameter empty for the default display.
-	\|~~GENE~~= ~~MAPK14, CSBP, CSBP1, CSBP2, MXI2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=9606 Homo sapiens])~~	+	-->
-	\|~~DOMAIN~~=	+	{{STRUCTURE_2pkj\| PDB=2pkj \| SCENE= }}
-	~~\|RELATEDENTRY=~~	+
-	\|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2pkj FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2pkj OCA], [http://www.ebi.ac.uk/pdbsum/2pkj PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2pkj RCSB]</span>	+
-	}}	+

	'''Crystal Structure of F169A p38 Kinase'''		'''Crystal Structure of F169A p38 Kinase'''
Line 30:		Line 27:
	[[Category: Namboodiri, H V.M.]]		[[Category: Namboodiri, H V.M.]]
	[[Category: Springman, E B.]]		[[Category: Springman, E B.]]
-	[[Category: ~~protein~~ kinase]]	+	[[Category: Protein kinase]]
-		+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 13:18:19 2008''
-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on ~~Mon Mar 31 04~~:37:11 2008''	+

OCA at 01:37, 31 March 2008

OCA — Mon, 31 Mar 2008 01:37:13 GMT

←Older revision		Revision as of 01:37, 31 March 2008
Line 5:		Line 5:
	\|SITE=		\|SITE=
	\|LIGAND= <scene name='pdbligand=BOG:B-OCTYLGLUCOSIDE'>BOG</scene>		\|LIGAND= <scene name='pdbligand=BOG:B-OCTYLGLUCOSIDE'>BOG</scene>
-	\|ACTIVITY= [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24]	+	\|ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] </span>
	\|GENE= MAPK14, CSBP, CSBP1, CSBP2, MXI2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=9606 Homo sapiens])		\|GENE= MAPK14, CSBP, CSBP1, CSBP2, MXI2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=9606 Homo sapiens])
		+	\|DOMAIN=
		+	\|RELATEDENTRY=
		+	\|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2pkj FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2pkj OCA], [http://www.ebi.ac.uk/pdbsum/2pkj PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2pkj RCSB]</span>
	}}		}}

Line 27:		Line 30:
	[[Category: Namboodiri, H V.M.]]		[[Category: Namboodiri, H V.M.]]
	[[Category: Springman, E B.]]		[[Category: Springman, E B.]]
-	[[Category: BOG]]
	[[Category: protein kinase]]		[[Category: protein kinase]]

-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on ~~Thu~~ Mar ~~20 18~~:14:09 2008''	+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 04:37:11 2008''

OCA at 16:14, 20 March 2008

OCA — Thu, 20 Mar 2008 16:14:09 GMT

←Older revision		Revision as of 16:14, 20 March 2008
Line 1:		Line 1:
-	[[Image:2pkj.jpg\|left\|200px]]~~<br /><applet load~~="2pkj~~" size~~="350~~" color~~=~~"white" frame="true" align~~=~~"right" spinBox="true"~~	+	[[Image:2pkj.jpg\|left\|200px]]
-	~~caption="~~2pkj, resolution 2.000Å" />	+
-	'''Crystal Structure of F169A p38 Kinase'''~~<br />~~	+	{{Structure
		+	\|PDB= 2pkj \|SIZE=350\|CAPTION= <scene name='initialview01'>2pkj</scene>, resolution 2.000Å
		+	\|SITE=
		+	\|LIGAND= <scene name='pdbligand=BOG:B-OCTYLGLUCOSIDE'>BOG</scene>
		+	\|ACTIVITY= [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24]
		+	\|GENE= MAPK14, CSBP, CSBP1, CSBP2, MXI2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=9606 Homo sapiens])
		+	}}
		+
		+	'''Crystal Structure of F169A p38 Kinase'''
		+

	==Overview==		==Overview==
Line 7:		Line 16:

	==About this Structure==		==About this Structure==
-	2PKJ is a [~~http://en.wikipedia.org/wiki/Single_protein~~ Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] ~~with <scene name='pdbligand=BOG:'>BOG</scene> as [http://en~~.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].	+	2PKJ is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].

	==Reference==		==Reference==
-	Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state., Bukhtiyarova M, Karpusas M, Northrop K, Namboodiri HV, Springman EB, Biochemistry. 2007 May 15;46(19):5687-96. Epub 2007 Apr 19. PMID:[http://~~ispc~~.~~weizmann~~.ac.~~il/~~/~~pmbin~~/~~getpm?pmid=~~17441692 17441692]	+	Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state., Bukhtiyarova M, Karpusas M, Northrop K, Namboodiri HV, Springman EB, Biochemistry. 2007 May 15;46(19):5687-96. Epub 2007 Apr 19. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/17441692 17441692]
	[[Category: Homo sapiens]]		[[Category: Homo sapiens]]
	[[Category: Mitogen-activated protein kinase]]		[[Category: Mitogen-activated protein kinase]]
Line 21:		Line 30:
	[[Category: protein kinase]]		[[Category: protein kinase]]

-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu ~~Feb 21~~ 18:30:26 2008''	+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 18:14:09 2008''

OCA at 16:30, 21 February 2008

OCA — Thu, 21 Feb 2008 16:30:27 GMT

←Older revision		Revision as of 16:30, 21 February 2008
Line 4:		Line 4:

	==Overview==		==Overview==
-	In order to study the role of Phe169 in p38alpha MAP kinase structure and, function, wild-type p38alpha and five p38alpha DFG motif mutants were, examined in vitro for phosphorylation by MKK6, kinase activity toward ATF2, substrate, thermal stability, and X-ray crystal structure. All six, p38alpha variants were efficiently phosphorylated by MKK6. However, only, one activated p38alpha mutant (F169Y) possessed measurable kinase activity, (1% compared to wild-type). The loss of kinase activity among the DFG, mutants may result from an inability to correctly position Asp168 in the, activated form of p38alpha. Two mutations significantly increased the, thermal stability of p38alpha (F169A DeltaTm = 1.3 degrees C and D168G, DeltaTm = 3.8 degrees C), and two mutations significantly decreased the, stability of p38alpha (F169R DeltaTm = -3.2 degrees C and F169G DeltaTm =, -4.7 degrees C). Interestingly, X-ray crystal structures of two thermally, destabilized p38alpha-F169R and p38alpha-F169G mutants revealed a DFG-OUT, conformation in the absence of an inhibitor molecule. This DFG-OUT, conformation, termed alpha-DFG-OUT, is different from the ones previously, identified in p38alpha crystal structures with bound inhibitors and, postulated from high-temperature molecular dynamics simulations. Taken, together, these results indicate that Phe169 is optimized for p38alpha, functional activity and structural dynamics, rather than for structural, stability. The alpha-DFG-OUT conformation observed for p38alpha-F169R and, p38alpha-F169G may represent a naturally occurring intermediate state of, p38alpha that provides access for binding of allosteric inhibitors. A, model of the local forces driving the DFG IN-OUT transition in p38alpha is, proposed.	+	In order to study the role of Phe169 in p38alpha MAP kinase structure and function, wild-type p38alpha and five p38alpha DFG motif mutants were examined in vitro for phosphorylation by MKK6, kinase activity toward ATF2 substrate, thermal stability, and X-ray crystal structure. All six p38alpha variants were efficiently phosphorylated by MKK6. However, only one activated p38alpha mutant (F169Y) possessed measurable kinase activity (1% compared to wild-type). The loss of kinase activity among the DFG mutants may result from an inability to correctly position Asp168 in the activated form of p38alpha. Two mutations significantly increased the thermal stability of p38alpha (F169A DeltaTm = 1.3 degrees C and D168G DeltaTm = 3.8 degrees C), and two mutations significantly decreased the stability of p38alpha (F169R DeltaTm = -3.2 degrees C and F169G DeltaTm = -4.7 degrees C). Interestingly, X-ray crystal structures of two thermally destabilized p38alpha-F169R and p38alpha-F169G mutants revealed a DFG-OUT conformation in the absence of an inhibitor molecule. This DFG-OUT conformation, termed alpha-DFG-OUT, is different from the ones previously identified in p38alpha crystal structures with bound inhibitors and postulated from high-temperature molecular dynamics simulations. Taken together, these results indicate that Phe169 is optimized for p38alpha functional activity and structural dynamics, rather than for structural stability. The alpha-DFG-OUT conformation observed for p38alpha-F169R and p38alpha-F169G may represent a naturally occurring intermediate state of p38alpha that provides access for binding of allosteric inhibitors. A model of the local forces driving the DFG IN-OUT transition in p38alpha is proposed.

	==About this Structure==		==About this Structure==
Line 16:		Line 16:
	[[Category: Bukhtiyarova, M.]]		[[Category: Bukhtiyarova, M.]]
	[[Category: Karpusas, M.]]		[[Category: Karpusas, M.]]
-	[[Category: Namboodiri, H.V.M.]]	+	[[Category: Namboodiri, H V.M.]]
-	[[Category: Springman, E.B.]]	+	[[Category: Springman, E B.]]
	[[Category: BOG]]		[[Category: BOG]]
	[[Category: protein kinase]]		[[Category: protein kinase]]

-	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on ~~Wed Jan 23 14~~:56:10 2008''	+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:30:26 2008''

OCA at 12:56, 23 January 2008

OCA — Wed, 23 Jan 2008 12:56:10 GMT

←Older revision		Revision as of 12:56, 23 January 2008
Line 1:		Line 1:
-	[[Image:2pkj.~~gif~~\|left\|200px]]<br />	+	[[Image:2pkj.jpg\|left\|200px]]<br /><applet load="2pkj" size="350" color="white" frame="true" align="right" spinBox="true"
-	<applet load="2pkj" size="~~450~~" color="white" frame="true" align="right" spinBox="true"	+
	caption="2pkj, resolution 2.000Å" />		caption="2pkj, resolution 2.000Å" />
	'''Crystal Structure of F169A p38 Kinase'''<br />		'''Crystal Structure of F169A p38 Kinase'''<br />
Line 8:		Line 7:

	==About this Structure==		==About this Structure==
-	2PKJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with BOG as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].	+	2PKJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=BOG:'>BOG</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].

	==Reference==		==Reference==
Line 22:		Line 21:
	[[Category: protein kinase]]		[[Category: protein kinase]]

-	''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Mon Nov 12~~ 23:24:~~41 2007~~''	+	''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 14:56:10 2008''

OCA: New page: 200px
'''Crystal Structure ...
OCA — Mon, 12 Nov 2007 21:18:13 GMT

New page: 200px<br /> <applet load="2pkj" size="450" color="white" frame="true" align="right" spinBox="true" caption="2pkj, resolution 2.000Å" /> '''Crystal Structure ...

New page
[[Image:2pkj.gif|left|200px]]<br />
<applet load="2pkj" size="450" color="white" frame="true" align="right" spinBox="true"
caption="2pkj, resolution 2.000Å" />
'''Crystal Structure of F169A p38 Kinase'''<br />

==Overview==
In order to study the role of Phe169 in p38alpha MAP kinase structure and, function, wild-type p38alpha and five p38alpha DFG motif mutants were, examined in vitro for phosphorylation by MKK6, kinase activity toward ATF2, substrate, thermal stability, and X-ray crystal structure. All six, p38alpha variants were efficiently phosphorylated by MKK6. However, only, one activated p38alpha mutant (F169Y) possessed measurable kinase activity, (1% compared to wild-type). The loss of kinase activity among the DFG, mutants may result from an inability to correctly position Asp168 in the, activated form of p38alpha. Two mutations significantly increased the, thermal stability of p38alpha (F169A DeltaTm = 1.3 degrees C and D168G, DeltaTm = 3.8 degrees C), and two mutations significantly decreased the, stability of p38alpha (F169R DeltaTm = -3.2 degrees C and F169G DeltaTm =, -4.7 degrees C). Interestingly, X-ray crystal structures of two thermally, destabilized p38alpha-F169R and p38alpha-F169G mutants revealed a DFG-OUT, conformation in the absence of an inhibitor molecule. This DFG-OUT, conformation, termed alpha-DFG-OUT, is different from the ones previously, identified in p38alpha crystal structures with bound inhibitors and, postulated from high-temperature molecular dynamics simulations. Taken, together, these results indicate that Phe169 is optimized for p38alpha, functional activity and structural dynamics, rather than for structural, stability. The alpha-DFG-OUT conformation observed for p38alpha-F169R and, p38alpha-F169G may represent a naturally occurring intermediate state of, p38alpha that provides access for binding of allosteric inhibitors. A, model of the local forces driving the DFG IN-OUT transition in p38alpha is, proposed.

==About this Structure==
2PKJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with BOG as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Mitogen-activated_protein_kinase Mitogen-activated protein kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.24 2.7.11.24] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2PKJ OCA].

==Reference==
Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state., Bukhtiyarova M, Karpusas M, Northrop K, Namboodiri HV, Springman EB, Biochemistry. 2007 May 15;46(19):5687-96. Epub 2007 Apr 19. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17441692 17441692]
[[Category: Homo sapiens]]
[[Category: Mitogen-activated protein kinase]]
[[Category: Single protein]]
[[Category: Bukhtiyarova, M.]]
[[Category: Karpusas, M.]]
[[Category: Namboodiri, H.V.M.]]
[[Category: Springman, E.B.]]
[[Category: BOG]]
[[Category: protein kinase]]

''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 23:24:41 2007''