Walker‐A threonine couples nucleotide occupancy with the chaperone activity of the AAA+ ATPase ClpB

Nagy, Mária; Wu, Hui-Chuan; Z, Liu; Kędzierska‐Mieszkowska, Sabina; Żółkiewski, Michał

doi:10.1002/pro.36

Cited by 33 publications

(40 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The reason for this discrepancy might be the presence of KCl in our experiments, which is known to modulate the population of functional protein oligomers (19,51). As recently reported (36), the lack of a quantitative description of the properties of each of the 12 available nucleotide binding sites and of the inter-and intrasubunit signaling that might communicate them makes difficult the use of an appropriate binding model to fit the experimental data. The model used herein considers ligand binding to equivalent and independent NBD2 sites.…”

Section: Discussionmentioning

confidence: 99%

“…and ATP␥S binding to WT ClpB (36). As compared with ATP␥S, ATP binding to T1T2 and N1T2 results in an ϳ2-fold reduction in enthalpy and a favorable entropy change, suggesting that the analog does not fully mimic the effect of ATP.…”

Section: Table 2 Isothermal Titration Calorimetry Analysis Of Nucleotmentioning

confidence: 98%

“…As previously discussed for ClpB (36) and other related ATPases (37), data analysis is not straightforward due to the presence of 12 different nucleotide binding sites that might have distinct binding properties and might display cooperativity as a result of complex allosteric interactions between different binding sites. Although the value of the apparent thermodynamic parameters of nucleotide binding depends on the binding model used to analyze the data (dependent or independent binding sites, positive and/or negative cooperativity), the stoichiometry of the binding reaction can be estimated from titration experiments.…”

Section: The Nucleotide Binding Domains Of Clpb Have Differentmentioning

confidence: 99%

See 2 more Smart Citations

Allosteric Communication between the Nucleotide Binding Domains of Caseinolytic Peptidase B

Fernández-Higuero

Acebrón

Taneva

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

ClpB is a hexameric chaperone that solubilizes and reactivates protein aggregates in cooperation with the Hsp70/DnaK chaperone system. Each of the identical protein monomers contains two nucleotide binding domains (NBD), whose ATPase activity must be coupled to exert on the substrate the mechanical work required for its reactivation. However, how communication between these sites occurs is at present poorly understood. We have studied herein the affinity of each of the NBDs for nucleotides in WT ClpB and protein variants in which one or both sites are mutated to selectively impair nucleotide binding or hydrolysis. Our data show that the affinity of NBD2 for nucleotides (K d ‫؍‬ 3-7 M) is significantly higher than that of NBD1. Interestingly, the affinity of NBD1 depends on nucleotide binding to NBD2. Binding of ATP, but not ADP, to NBD2 increases the affinity of NBD1 (the K d decreases from ≈160 -300 to 50 -60 M) for the corresponding nucleotide. Moreover, filling of the NBD2 ring with ATP allows the cooperative binding of this nucleotide and substrates to the NBD1 ring. Data also suggest that a minimum of four subunits cooperate to bind and reactivate two different aggregated protein substrates.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Table 2 Isothermal Titration Calorimetry Analysis Of Nucleotmentioning

confidence: 98%

Section: The Nucleotide Binding Domains Of Clpb Have Differentmentioning

confidence: 99%

See 1 more Smart Citation

Allosteric Communication between the Nucleotide Binding Domains of Caseinolytic Peptidase B

Fernández-Higuero

Acebrón

Taneva

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…DYT1 dystonia | nuclear envelope | ATPase | Torsin T orsin ATPases belong to the AAA+ (ATPase associated with a variety of cellular activities) superfamily of ATPases (1) but are unusual in that they lack conserved catalytic residues that are typically found in related ATPases (2,3). Accordingly, Torsins do not display ATPase activity unless they are engaged by their regulatory cofactors lamina-associated polypeptide 1 (LAP1) or luminal domain-like LAP1 (LULL1) (4), which are type II transmembrane proteins residing in the nuclear envelope and endoplasmic reticulum (ER) (5,6).…”

mentioning

confidence: 99%

The mechanism of Torsin ATPase activation

Brown

Zhao

Chase

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

142

View full text Add to dashboard Cite

Torsins are membrane-associated ATPases whose activity is dependent on two activating cofactors, lamina-associated polypeptide 1 (LAP1) and luminal domain-like LAP1 (LULL1). The mechanism by which these cofactors regulate Torsin activity has so far remained elusive. In this study, we identify a conserved domain in these activators that is predicted to adopt a fold resembling an AAA+ (ATPase associated with a variety of cellular activities) domain. Within these domains, a strictly conserved Arg residue present in both activating cofactors, but notably missing in Torsins, aligns with a key catalytic Arg found in AAA+ proteins. We demonstrate that cofactors and Torsins associate to form heterooligomeric assemblies with a defined Torsin-activator interface. In this arrangement, the highly conserved Arg residue present in either cofactor comes into close proximity with the nucleotide bound in the neighboring Torsin subunit. Because this invariant Arg is strictly required to stimulate Torsin ATPase activity but is dispensable for Torsin binding, we propose that LAP1 and LULL1 regulate Torsin ATPase activity through an active site complementation mechanism.DYT1 dystonia | nuclear envelope | ATPase | Torsin T orsin ATPases belong to the AAA+ (ATPase associated with a variety of cellular activities) superfamily of ATPases (1) but are unusual in that they lack conserved catalytic residues that are typically found in related ATPases (2, 3). Accordingly, Torsins do not display ATPase activity unless they are engaged by their regulatory cofactors lamina-associated polypeptide 1 (LAP1) or luminal domain-like LAP1 (LULL1) (4), which are type II transmembrane proteins residing in the nuclear envelope and endoplasmic reticulum (ER) (5, 6). This property stands in sharp contrast to the behavior of closely related Clp/Hsp100 ATPases, which display considerable basal ATPase activities that are moderately stimulated by their substrates (7,8). Although a number of Clp/Hsp100 proteins use distinct cofactors that confer substrate specificity to the typically hexameric ATPase ring, they operate via similar principles (9). Substrates ultimately engage the pore at the center of the oligomeric assembly. This narrow annulus is defined by pore loops that emanate from each subunit and harbor a conserved aromatic residue that defines the center of the pore (10). The energy of ATP hydrolysis is invested in threading the substrate through this axial channel, and the substrate is unfolded in the process (11).Much less is known about the mode of action of Torsin ATPases, although it is clear that regulation of Torsin activity is of vital importance in an organismal context. A mutation in TorsinA (TorA) causing the movement disorder primary dystonia (12) renders TorA unresponsive to its binding partners LAP1 and LULL1 (4), and a homozygous "knock-in" of this disease allele in mice causes a lethal phenotype, as does a TorA KO (13). A conditional deletion of TorA from the CNS in mice accurately replicates the symptoms of primary dystonia (14). In add...

show abstract

“…In some proteins, such as a subset of AAA+ ATPases, the conserved Thr/Ser of the Walker-A motif is replaced by Asn, yielding GX 4 GKN. Substitution of the conserved Thr in another AAA+ ATPase, ClpB, with Asn, resulted in an enzyme in which nucleotide-binding and activity were effectively uncoupled, demonstrating the functional importance of the conserved Thr in most AAA+ ATPases (Nagy et al, 2009 Protein kinases exhibit a conserved core structure of an Nand C-terminal domain, separated by a cleft. Nucleotidebinding involves primarily the N-terminal domain, involving interactions with residues from b-strands within this segment.…”

Section: +mentioning

confidence: 99%

ATP ‐binding Motifs

Matte

Delbaere

2010

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Adenosine 5′‐triphosphate (ATP) binds to a great number of proteins to elicit a wide variety of effects, including energy production and molecular signalling. Proteins have evolved different strategies to specifically recognize ATP, utilizing different ways of binding the phosphoryl moieties as well as the adenine base. The most common, conserved sequence and structural motif for binding ATP is the Walker‐A motif, or P‐loop, found in many different protein structural families. Greater variation in the sequence of the P‐loop is being recognized, as more ATP‐binding proteins are being structurally and functionally characterized. In contrast to the P‐loop, recognition of the adenine base often makes use of conserved structural motifs of main‐chain atoms via hydrogen‐bonding interactions, or side‐chains in stacking interactions, without a definitive amino acid sequence pattern. Key concepts: A major class of ATP‐binding proteins are those that contain a P‐loop or Walker‐A motif. P‐loops or glycine‐rich loops function by binding the phosphoryl groups of ATP. Several sequence variations on the Walker‐A motif are now known and have been functionally characterized. The Walker‐B motif contains a conserved acidic residue (Glu/Asp) that functions to bind directly or indirectly a metal ion important in catalysis. Adenine‐binding does not occur through specific sequence motifs, but rather uses a conserved pattern of polar and nonpolar interactions within a structural motif. Both main‐chain hydrogen bonding and aromatic residue stacking contribute to adenine‐binding by proteins.

show abstract

Walker‐A threonine couples nucleotide occupancy with the chaperone activity of the AAA+ ATPase ClpB

Cited by 33 publications

References 29 publications

Allosteric Communication between the Nucleotide Binding Domains of Caseinolytic Peptidase B

Allosteric Communication between the Nucleotide Binding Domains of Caseinolytic Peptidase B

The mechanism of Torsin ATPase activation

ATP ‐binding Motifs

Contact Info

Product

Resources

About