Substrate Binding Causes Movement in the ATP Binding Domain ofEscherichia coliAdenylate Kinase†

Biochemistry ◽  
1996 ◽  
Vol 35 (19) ◽  
pp. 6100-6106 ◽  
Author(s):  
Tim Bilderback ◽  
Tim Fulmer ◽  
William W. Mantulin ◽  
Michael Glaser
Keyword(s):  
Substrate Binding ◽  
Binding Domain ◽  
Atp Binding

scholarly journals Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase

2017 ◽  
Vol 37 (6) ◽  
Author(s):  
Jungsoon Lee ◽  
Nuri Sung ◽  
Lythou Yeo ◽  
Changsoo Chang ◽  
Sukyeong Lee ◽  
...  
Keyword(s):  
Protein Unfolding ◽  
Substrate Binding ◽  
Binding Domain ◽  
Atp Binding ◽  
Structural Determinants ◽  
Functional Studies ◽  
Loop 2 ◽  
Loop Conformation ◽  
Do So

The ring-forming Hsp104 ATPase cooperates with Hsp70 and Hsp40 molecular chaperones to rescue stress-damaged proteins from both amorphous and amyloid-forming aggregates. The ability to do so relies upon pore loops present in the first ATP-binding domain (AAA-1; loop-1 and loop-2 ) and in the second ATP-binding domain (AAA-2; loop-3) of Hsp104, which face the protein translocating channel and couple ATP-driven changes in pore loop conformation to substrate translocation. A hallmark of loop-1 and loop-3 is an invariable and mutational sensitive aromatic amino acid (Tyr257 and Tyr662) involved in substrate binding. However, the role of conserved aliphatic residues (Lys256, Lys258, and Val663) flanking the pore loop tyrosines, and the function of loop-2 in protein disaggregation has not been investigated. Here we present the crystal structure of an N-terminal fragment of Saccharomyces cerevisiae Hsp104 exhibiting molecular interactions involving both AAA-1 pore loops, which resemble contacts with bound substrate. Corroborated by biochemical experiments and functional studies in yeast, we show that aliphatic residues flanking Tyr257 and Tyr662 are equally important for substrate interaction, and abolish Hsp104 function when mutated to glycine. Unexpectedly, we find that loop-2 is sensitive to aspartate substitutions that impair Hsp104 function and abolish protein disaggregation when loop-2 is replaced by four aspartate residues. Our observations suggest that Hsp104 pore loops have non-overlapping functions in protein disaggregation and together coordinate substrate binding, unfolding, and translocation through the Hsp104 hexamer.

A Three-Dimensional Model for the Substrate Binding Domain of the Multidrug ATP Binding Cassette Transporter LmrA

2004 ◽  
Vol 66 (5) ◽  
pp. 1169-1179 ◽  
Author(s):  
Gerhard F. Ecker ◽  
Karin Pleban ◽  
Stephan Kopp ◽  
Edina Csaszar ◽  
Gerrit J. Poelarends ◽  
...  
Keyword(s):  
Substrate Binding ◽  
Three Dimensional ◽  
Binding Domain ◽  
Atp Binding ◽  
Dimensional Model ◽  
Three Dimensional Model ◽  
Atp Binding Cassette Transporter ◽  
Substrate Binding Domain ◽  
Atp Binding Cassette

scholarly journals Mutagenic Analysis of the Putative ABCC6 Substrate-Binding Cavity Using a New Homology Model

2021 ◽  
Vol 22 (13) ◽  
pp. 6910
Author(s):  
Flora Szeri ◽  
Valentina Corradi ◽  
Fatemeh Niaziorimi ◽  
Sylvia Donnelly ◽  
Gwenaëlle Conseil ◽  
...  
Keyword(s):  
Binding Site ◽  
Efflux Pump ◽  
Substrate Binding ◽  
Functional Characterization ◽  
Atp Release ◽  
Homology Model ◽  
Atp Binding ◽  
Atp Binding Site ◽  
Binding Cavity ◽  
Atp Efflux

Inactivating mutations in ABCC6 underlie the rare hereditary mineralization disorder pseudoxanthoma elasticum. ABCC6 is an ATP-binding cassette (ABC) integral membrane protein that mediates the release of ATP from hepatocytes into the bloodstream. The released ATP is extracellularly converted into pyrophosphate, a key mineralization inhibitor. Although ABCC6 is firmly linked to cellular ATP release, the molecular details of ABCC6-mediated ATP release remain elusive. Most of the currently available data support the hypothesis that ABCC6 is an ATP-dependent ATP efflux pump, an un-precedented function for an ABC transporter. This hypothesis implies the presence of an ATP-binding site in the substrate-binding cavity of ABCC6. We performed an extensive mutagenesis study using a new homology model based on recently published structures of its close homolog, bovine Abcc1, to characterize the substrate-binding cavity of ABCC6. Leukotriene C4 (LTC4), is a high-affinity substrate of ABCC1. We mutagenized fourteen amino acid residues in the rat ortholog of ABCC6, rAbcc6, that corresponded to the residues in ABCC1 found in the LTC4 binding cavity. Our functional characterization revealed that most of the amino acids in rAbcc6 corresponding to those found in the LTC4 binding pocket in bovine Abcc1 are not critical for ATP efflux. We conclude that the putative ATP binding site in the substrate-binding cavity of ABCC6/rAbcc6 is distinct from the bovine Abcc1 LTC4-binding site.

scholarly journals Entropic Inhibition: How the Activity of a AAA+ Machine Is Modulated by Its Substrate-Binding Domain

2021 ◽  
Author(s):  
Marija Iljina ◽  
Hisham Mazal ◽  
Pierre Goloubinoff ◽  
Inbal Riven ◽  
Gilad Haran
Keyword(s):  
Substrate Binding ◽  
Binding Domain ◽  
Substrate Binding Domain

Phosphate Binding to Isolated Chloroplast Coupling Factor (CF1)

1988 ◽  
Vol 43 (3-4) ◽  
pp. 213-218 ◽  
Author(s):  
Bernhard Huchzermeyer
Keyword(s):  
Atpase Activity ◽  
Binding Site ◽  
Inorganic Phosphate ◽  
Coupling Factor ◽  
Binding Domain ◽  
Atp Binding ◽  
Phosphate Binding ◽  
Chloroplast Coupling Factor ◽  
Single Binding Site ◽  
Site Binding

A single binding site for phosphate was found on isolated chloroplast coupling factor in the absence of nucleotides. In our experiments the phosphate binding site showed a Kd of 170 μᴍ. We did not observe any differences whether the ATPase activity of CF] had been activated or not. If the enzyme was incubated with [γ-32P]ATP the amount of 32P bound per CF1 depended on the pretreatment of the enzyme: In the presence of ADP no ATP or phosphate was bound to CF,. After activation of ATPase activity one mol of ATP per mol CF, was rapidly bound and hydrolyzed while there was a slowly occurring binding of another phosphate without concomitant nucleotide binding. We conclude that there are two different types of phosphate binding observed in our experiments: 1) Inorganic phosphate can be bound by one catalytic site per mol of CF1 2) The γ-phosphate of ATP is able to bind to an ATP binding domain of the enzyme if this domain can exchange substrates with the incubation medium. This ATP binding domain appears to differ from the site binding inorganic phosphate, because at least a portion of the coupling factor contains more than one labelled phosphate during our ATPase tests.

scholarly journals Flexible Linkers Leash the Substrate Binding Domain of SspB to a Peptide Module that Stabilizes Delivery Complexes with the AAA+ ClpXP Protease

2003 ◽  
Vol 12 (2) ◽  
pp. 355-363 ◽  
Author(s):  
David A Wah ◽  
Igor Levchenko ◽  
Gabrielle E Rieckhof ◽  
Daniel N Bolon ◽  
Tania A Baker ◽  
...  
Keyword(s):  
Substrate Binding ◽  
Binding Domain ◽  
Substrate Binding Domain

scholarly journals An atomistic view of Hsp70 allosteric crosstalk: from the nucleotide to the substrate binding domain and back

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Federica Chiappori ◽  
Ivan Merelli ◽  
Luciano Milanesi ◽  
Giorgio Colombo ◽  
Giulia Morra
Keyword(s):  
Substrate Binding ◽  
Binding Domain ◽  
Substrate Binding Domain

scholarly journals Dissection of the ATP-binding Domain of the Chaperone hsc70 for Interaction with the Cofactor Hap46

2000 ◽  
Vol 276 (13) ◽  
pp. 10178-10184 ◽  
Author(s):  
Gabriele Petersen ◽  
Christian Hahn ◽  
Ulrich Gehring
Keyword(s):  
Binding Domain ◽  
Atp Binding
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