4g4e

From Proteopedia

(Difference between revisions)

Current revision

Crystal structure of the L88A mutant of HslV from Escherichia coli

Structural highlights

4g4e is a 12 chain structure with sequence from Escherichia coli K-12. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 2.888Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

HSLV_ECOLI Protease subunit of a proteasome-like degradation complex believed to be a general protein degrading machinery. The complex has been shown to be involved in the specific degradation of heat shock induced transcription factors such as RpoH and SulA. In addition, small hydrophobic peptides are also hydrolyzed by HslV. HslV has weak protease activity even in the absence of HslU, but this activity is induced more than 100-fold in the presence of HslU. HslU recognizes protein substrates and unfolds these before guiding them to HslV for hydrolysis. HslV is not believed to degrade folded proteins.^[1] ^[2] ^[3] ^[4] ^[5] ^[6] ^[7]

Publication Abstract from PubMed

In all cells, ATP-dependent proteases play central roles in the controlled degradation of short-lived regulatory or misfolded proteins. A hallmark of these enzymes is that proteolytic active sites are sequestered within a compartmentalized space, which is accessible to substrates only when they are fed into the cavity by protein-unfolding ATPases. HslVU is a prototype of such enzymes, consisting of the hexameric HslU ATPase and the dodecameric HslV protease. HslV forms a barrel-shaped proteolytic chamber with two constricted axial pores. Here, we report that structural alterations of HslV's pore motif dramatically affect the proteolytic activities of both HslV and HslVU complexes. Mutations of a conserved pore residue in HslV (Leu88 to Ala, Gly, or Ser) led to a tighter binding between HslV and HslU and a dramatic stimulation of both the proteolytic and ATPase activities. Furthermore, the HslV mutants alone showed a marked increase of basal hydrolytic activities toward small peptides and unstructured proteins. A synthetic peptide of the HslU C-terminal tail further stimulated the proteolytic activities of these mutants, even allowing degradation of certain folded proteins in the absence of HslU. Moreover, expression of the L88A mutant in Escherichia coli inhibited cell growth, suggesting that HslV pore mutations dysregulate the protease through relaxing the pore constriction, which normally prevents essential cellular proteins from random degradation. Consistent with these observations, an X-ray crystal structure shows that the pore loop of L88A-HslV is largely disordered. Collectively, these results suggest that substrate degradation by HslV is controlled by gating of its pores.

Structural Alteration in the Pore Motif of the Bacterial 20S Proteasome Homolog HslV Leads to Uncontrolled Protein Degradation.,Park E, Lee JW, Yoo HM, Ha BH, An JY, Jeon YJ, Seol JH, Eom SH, Chung CH J Mol Biol. 2013 May 21. pii: S0022-2836(13)00310-0. doi:, 10.1016/j.jmb.2013.05.011. PMID:23707406^[8]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Yoo SJ, Seol JH, Shin DH, Rohrwild M, Kang MS, Tanaka K, Goldberg AL, Chung CH. Purification and characterization of the heat shock proteins HslV and HslU that form a new ATP-dependent protease in Escherichia coli. J Biol Chem. 1996 Jun 14;271(24):14035-40. PMID:8662828
↑ Rohrwild M, Coux O, Huang HC, Moerschell RP, Yoo SJ, Seol JH, Chung CH, Goldberg AL. HslV-HslU: A novel ATP-dependent protease complex in Escherichia coli related to the eukaryotic proteasome. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5808-13. PMID:8650174
↑ Seol JH, Yoo SJ, Shin DH, Shim YK, Kang MS, Goldberg AL, Chung CH. The heat-shock protein HslVU from Escherichia coli is a protein-activated ATPase as well as an ATP-dependent proteinase. Eur J Biochem. 1997 Aug 1;247(3):1143-50. PMID:9288941
↑ Kanemori M, Nishihara K, Yanagi H, Yura T. Synergistic roles of HslVU and other ATP-dependent proteases in controlling in vivo turnover of sigma32 and abnormal proteins in Escherichia coli. J Bacteriol. 1997 Dec;179(23):7219-25. PMID:9393683
↑ Seong IS, Oh JY, Yoo SJ, Seol JH, Chung CH. ATP-dependent degradation of SulA, a cell division inhibitor, by the HslVU protease in Escherichia coli. FEBS Lett. 1999 Jul 30;456(1):211-4. PMID:10452560
↑ Kanemori M, Yanagi H, Yura T. Marked instability of the sigma(32) heat shock transcription factor at high temperature. Implications for heat shock regulation. J Biol Chem. 1999 Jul 30;274(31):22002-7. PMID:10419524
↑ Burton RE, Baker TA, Sauer RT. Nucleotide-dependent substrate recognition by the AAA+ HslUV protease. Nat Struct Mol Biol. 2005 Mar;12(3):245-51. Epub 2005 Feb 6. PMID:15696175 doi:10.1038/nsmb898
↑ Park E, Lee JW, Yoo HM, Ha BH, An JY, Jeon YJ, Seol JH, Eom SH, Chung CH. Structural Alteration in the Pore Motif of the Bacterial 20S Proteasome Homolog HslV Leads to Uncontrolled Protein Degradation. J Mol Biol. 2013 May 21. pii: S0022-2836(13)00310-0. doi:, 10.1016/j.jmb.2013.05.011. PMID:23707406 doi:10.1016/j.jmb.2013.05.011

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 See Also
5 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4g4e"

@@ Line 1: / Line 1: @@
-{{STRUCTURE_4g4e|  PDB=4g4e  |  SCENE=  }}
-===Crystal structure of the L88A mutant of HslV from Escherichia coli===
-{{ABSTRACT_PUBMED_23707406}}
-==Function==
+==Crystal structure of the L88A mutant of HslV from Escherichia coli==
-[[http://www.uniprot.org/uniprot/HSLV_ECOLI HSLV_ECOLI]] Protease subunit of a proteasome-like degradation complex believed to be a general protein degrading machinery. The complex has been shown to be involved in the specific degradation of heat shock induced transcription factors such as RpoH and SulA. In addition, small hydrophobic peptides are also hydrolyzed by HslV. HslV has weak protease activity even in the absence of HslU, but this activity is induced more than 100-fold in the presence of HslU. HslU recognizes protein substrates and unfolds these before guiding them to HslV for hydrolysis. HslV is not believed to degrade folded proteins.<ref>PMID:8662828</ref> <ref>PMID:8650174</ref> <ref>PMID:9288941</ref> <ref>PMID:9393683</ref> <ref>PMID:10452560</ref> <ref>PMID:10419524</ref> <ref>PMID:15696175</ref>
+<StructureSection load='4g4e' size='340' side='right'caption='[[4g4e]], [[Resolution|resolution]] 2.89&Aring;' scene=''>
+== Structural highlights ==
+<table><tr><td colspan='2'>[[4g4e]] is a 12 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4G4E OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4G4E FirstGlance]. <br>
+</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.888&#8491;</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=4g4e FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4g4e OCA], [https://pdbe.org/4g4e PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4g4e RCSB], [https://www.ebi.ac.uk/pdbsum/4g4e PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4g4e ProSAT]</span></td></tr>
+</table>
+== Function ==
+[https://www.uniprot.org/uniprot/HSLV_ECOLI HSLV_ECOLI] Protease subunit of a proteasome-like degradation complex believed to be a general protein degrading machinery. The complex has been shown to be involved in the specific degradation of heat shock induced transcription factors such as RpoH and SulA. In addition, small hydrophobic peptides are also hydrolyzed by HslV. HslV has weak protease activity even in the absence of HslU, but this activity is induced more than 100-fold in the presence of HslU. HslU recognizes protein substrates and unfolds these before guiding them to HslV for hydrolysis. HslV is not believed to degrade folded proteins.<ref>PMID:8662828</ref> <ref>PMID:8650174</ref> <ref>PMID:9288941</ref> <ref>PMID:9393683</ref> <ref>PMID:10452560</ref> <ref>PMID:10419524</ref> <ref>PMID:15696175</ref>
+<div style="background-color:#fffaf0;">
+== Publication Abstract from PubMed ==
+In all cells, ATP-dependent proteases play central roles in the controlled degradation of short-lived regulatory or misfolded proteins. A hallmark of these enzymes is that proteolytic active sites are sequestered within a compartmentalized space, which is accessible to substrates only when they are fed into the cavity by protein-unfolding ATPases. HslVU is a prototype of such enzymes, consisting of the hexameric HslU ATPase and the dodecameric HslV protease. HslV forms a barrel-shaped proteolytic chamber with two constricted axial pores. Here, we report that structural alterations of HslV's pore motif dramatically affect the proteolytic activities of both HslV and HslVU complexes. Mutations of a conserved pore residue in HslV (Leu88 to Ala, Gly, or Ser) led to a tighter binding between HslV and HslU and a dramatic stimulation of both the proteolytic and ATPase activities. Furthermore, the HslV mutants alone showed a marked increase of basal hydrolytic activities toward small peptides and unstructured proteins. A synthetic peptide of the HslU C-terminal tail further stimulated the proteolytic activities of these mutants, even allowing degradation of certain folded proteins in the absence of HslU. Moreover, expression of the L88A mutant in Escherichia coli inhibited cell growth, suggesting that HslV pore mutations dysregulate the protease through relaxing the pore constriction, which normally prevents essential cellular proteins from random degradation. Consistent with these observations, an X-ray crystal structure shows that the pore loop of L88A-HslV is largely disordered. Collectively, these results suggest that substrate degradation by HslV is controlled by gating of its pores.
-==About this Structure==
+Structural Alteration in the Pore Motif of the Bacterial 20S Proteasome Homolog HslV Leads to Uncontrolled Protein Degradation.,Park E, Lee JW, Yoo HM, Ha BH, An JY, Jeon YJ, Seol JH, Eom SH, Chung CH J Mol Biol. 2013 May 21. pii: S0022-2836(13)00310-0. doi:, 10.1016/j.jmb.2013.05.011. PMID:23707406<ref>PMID:23707406</ref>
-[[4g4e]] is a 12 chain structure with sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_k-12 Escherichia coli k-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4G4E OCA].
-==Reference==
+From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
-<ref group="xtra">PMID:023707406</ref><references group="xtra"/><references/>
+</div>
-[[Category: Escherichia coli k-12]]
+<div class="pdbe-citations 4g4e" style="background-color:#fffaf0;"></div>
-[[Category: HslU--HslV peptidase]]
-[[Category: An, J Y.]]
+==See Also==
-[[Category: Chung, C H.]]
+*[[Heat Shock Protein structures|Heat Shock Protein structures]]
-[[Category: Eom, S H.]]
+== References ==
-[[Category: Ha, B H.]]
+<references/>
-[[Category: Jeon, Y J.]]
+__TOC__
-[[Category: Lee, J W.]]
+</StructureSection>
-[[Category: Park, E.]]
+[[Category: Escherichia coli K-12]]
-[[Category: Seol, J H.]]
+[[Category: Large Structures]]
-[[Category: Yoo, H M.]]
+[[Category: An JY]]
-[[Category: Atp-dependent protease]]
+[[Category: Chung CH]]
-[[Category: Hslu atpase]]
+[[Category: Eom SH]]
-[[Category: Hslv protease]]
+[[Category: Ha BH]]
-[[Category: Hslvu]]
+[[Category: Jeon YJ]]
-[[Category: Hydrolase]]
+[[Category: Lee JW]]
-[[Category: Pore motif]]
+[[Category: Park E]]
+[[Category: Seol JH]]
+[[Category: Yoo HM]]

4g4e

From Proteopedia

Current revision

Crystal structure of the L88A mutant of HslV from Escherichia coli

Structural highlights

Function

Publication Abstract from PubMed

See Also

References

Contents

Proteopedia Page Contributors and Editors (what is this?)

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