PMID- 24343376
OWN - NLM
STAT- MEDLINE
DCOM- 20141107
LR  - 20211021
IS  - 1433-4909 (Electronic)
IS  - 1431-0651 (Print)
IS  - 1431-0651 (Linking)
VI  - 18
IP  - 2
DP  - 2014 Mar
TI  - Structural and biochemical properties of an extreme 'salt-loving' proteasome 
      activating nucleotidase from the archaeon Haloferax volcanii.
PG  - 283-93
LID - 10.1007/s00792-013-0615-8 [doi]
AB  - In eukaryotes, the 26S proteasome degrades ubiquitinylated proteins in an 
      ATP-dependent manner. Archaea mediate a form of post-translational modification 
      of proteins termed sampylation that resembles ubiquitinylation. Sampylation was 
      identified in Haloferax volcanii, a moderate halophilic archaeon that synthesizes 
      homologs of 26S proteasome subunits including 20S core particles and regulatory 
      particle triple-A ATPases (Rpt)-like proteasome-associated nucleotidases (PAN-A/1 
      and PAN-B/2). To determine whether sampylated proteins associate with the Rpt 
      subunit homologs, PAN-A/1 was purified to homogeneity from Hfx. volcanii and 
      analyzed for its subunit stoichiometry, nucleotide-hydrolyzing activity and 
      binding to sampylated protein targets. PAN-A/1 was found to be associated as a 
      dodecamer (630 kDa) with a configuration in TEM suggesting a complex of two 
      stacked hexameric rings. PAN-A/1 had high affinity for ATP (K m of ~0.44 mM) and 
      hydrolyzed this nucleotide with a specific activity of 0.33 +/- 0.1 mumol Pi/h per 
      mg protein and maximum at 42  degrees C. PAN-A1 was stabilized by 2 M salt with a 
      decrease in activity at lower concentrations of salt that correlated with 
      dissociation of the dodecamer into trimers to monomers. Binding of PAN-A/1 to a 
      sampylated protein was demonstrated by modification of a far Western blotting 
      technique (derived from the standard Western blot method to detect 
      protein-protein interaction in vitro) for halophilic proteins. Overall, our 
      results support a model in which sampylated proteins associate with the PAN-A/1 
      AAA+ ATPase in proteasome-mediated proteolysis and/or protein remodeling and 
      provide a method for assay of halophilic protein-protein interactions.
FAU - Prunetti, Laurence
AU  - Prunetti L
AD  - Department of Microbiology and Cell Science, University of Florida, Gainesville, 
      FL, 32611-0700, USA, lprunetti@ufl.edu.
FAU - Reuter, Christopher J
AU  - Reuter CJ
FAU - Hepowit, Nathaniel L
AU  - Hepowit NL
FAU - Wu, Yifei
AU  - Wu Y
FAU - Barrueto, Luisa
AU  - Barrueto L
FAU - Miranda, Hugo V
AU  - Miranda HV
FAU - Kelly, Karen
AU  - Kelly K
FAU - Maupin-Furlow, Julie A
AU  - Maupin-Furlow JA
LA  - eng
GR  - R01 GM057498/GM/NIGMS NIH HHS/United States
GR  - R01 GM57498/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20131217
PL  - Germany
TA  - Extremophiles
JT  - Extremophiles : life under extreme conditions
JID - 9706854
RN  - 0 (Archaeal Proteins)
RN  - EC 3.1.3.- (Nucleotidases)
RN  - EC 3.1.3.31 (nucleotidase)
RN  - EC 3.4.25.1 (Proteasome Endopeptidase Complex)
SB  - IM
MH  - Archaeal Proteins/chemistry/*metabolism
MH  - Haloferax volcanii/*enzymology/physiology
MH  - Nucleotidases/chemistry/*metabolism
MH  - Osmolar Concentration
MH  - Proteasome Endopeptidase Complex/*metabolism
MH  - Protein Binding
MH  - Protein Multimerization
MH  - *Salt Tolerance
PMC - PMC3943764
MID - NIHMS551304
EDAT- 2013/12/18 06:00
MHDA- 2014/11/08 06:00
PMCR- 2015/03/01
CRDT- 2013/12/18 06:00
PHST- 2013/09/24 00:00 [received]
PHST- 2013/11/21 00:00 [accepted]
PHST- 2013/12/18 06:00 [entrez]
PHST- 2013/12/18 06:00 [pubmed]
PHST- 2014/11/08 06:00 [medline]
PHST- 2015/03/01 00:00 [pmc-release]
AID - 10.1007/s00792-013-0615-8 [doi]
PST - ppublish
SO  - Extremophiles. 2014 Mar;18(2):283-93. doi: 10.1007/s00792-013-0615-8. Epub 2013 
      Dec 17.