Structure and Mechanisms of F-Type ATP Synthases

2019 ◽  
Vol 88 (1) ◽  
pp. 515-549 ◽  
Author(s):  
Werner Kühlbrandt
Keyword(s):  
Molecular Mechanisms ◽  
Power Stroke ◽  
Hydrophobic Core ◽  
Joint Rotation ◽  
Γ Subunit ◽  
Mitochondrial Atp Synthase ◽  
Redox Switch ◽  
Rotary Catalysis ◽  
Unique Mechanism ◽  
Atp Synthases

F1Fo ATP synthases produce most of the ATP in the cell. F-type ATP synthases have been investigated for more than 50 years, but a full understanding of their molecular mechanisms has become possible only with the recent structures of complete, functionally competent complexes determined by electron cryo-microscopy (cryo-EM). High-resolution cryo-EM structures offer a wealth of unexpected new insights. The catalytic F1 head rotates with the central γ-subunit for the first part of each ATP-generating power stroke. Joint rotation is enabled by subunit δ/OSCP acting as a flexible hinge between F1 and the peripheral stalk. Subunit a conducts protons to and from the c-ring rotor through two conserved aqueous channels. The channels are separated by ∼6 Å in the hydrophobic core of Fo, resulting in a strong local field that generates torque to drive rotary catalysis in F1. The structure of the chloroplast F1Fo complex explains how ATPase activity is turned off at night by a redox switch. Structures of mitochondrial ATP synthase dimers indicate how they shape the inner membrane cristae. The new cryo-EM structures complete our picture of the ATP synthases and reveal the unique mechanism by which they transform an electrochemical membrane potential into biologically useful chemical energy.

scholarly journals Structural Basis for the C-Terminal Domain of Mycobacterium tuberculosis Ribosome Maturation Factor RimM to Bind Ribosomal Protein S19

Biomolecules ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 597
Author(s):  
Haoran Zhang ◽  
Qiuxiang Zhou ◽  
Chenyun Guo ◽  
Liubin Feng ◽  
Huilin Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Ribosomal Protein ◽  
Solution Structure ◽  
Molecular Mechanisms ◽  
Structural Model ◽  
Ribosomal Subunit ◽  
Structural Basis ◽  
Hydrophobic Core ◽  
Terminal Domain ◽  
Ribosomal Protein S19

Multidrug-resistant tuberculosis (TB) is a serious threat to public health, calling for the development of new anti-TB drugs. Chaperon protein RimM, involved in the assembly of ribosomal protein S19 into 30S ribosomal subunit during ribosome maturation, is a potential drug target for TB treatment. The C-terminal domain (CTD) of RimM is primarily responsible for binding S19. However, both the CTD structure of RimM from Mycobacterium tuberculosis (MtbRimMCTD) and the molecular mechanisms underlying MtbRimMCTD binding S19 remain elusive. Here, we report the solution structure, dynamics features of MtbRimMCTD, and its interaction with S19. MtbRimMCTD has a rigid hydrophobic core comprised of a relatively conservative six-strand β-barrel, tailed with a short α-helix and interspersed with flexible loops. Using several biophysical techniques including surface plasmon resonance (SPR) affinity assays, nuclear magnetic resonance (NMR) assays, and molecular docking, we established a structural model of the MtbRimMCTD–S19 complex and indicated that the β4-β5 loop and two nonconserved key residues (D105 and H129) significantly contributed to the unique pattern of MtbRimMCTD binding S19, which might be implicated in a form of orthogonality for species-dependent RimM–S19 interaction. Our study provides the structural basis for MtbRimMCTD binding S19 and is beneficial to the further exploration of MtbRimM as a potential target for the development of new anti-TB drugs.

Rat mitochondrial ATP synthase ATP5G3: cloning and upregulation in pancreas after chronic ethanol feeding

2001 ◽  
Vol 6 (2) ◽  
pp. 91-98 ◽  
Author(s):  
HA-SHENG LI ◽  
JI-YING ZHANG ◽  
BRYAN S. THOMPSON ◽  
XIAO-YING DENG ◽  
MICHAEL E. FORD ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Cellular Response ◽  
Molecular Mechanisms ◽  
Metabolic Stress ◽  
Nuclear Gene ◽  
Chronic Ethanol ◽  
Ethanol Ingestion ◽  
Pancreatic Acinar Cells ◽  
Mitochondrial Atp Synthase ◽  
Ethanol Feeding

Individuals with chronic excessive alcohol ingestion are put at the risk of acute and chronic pancreatitis. Underlying molecular mechanisms are unknown. Differential gene expression in the pancreas was profiled using mRNA differential display by comparison between control and ethanol-consuming rats. Male Wistar rats were fed with diets containing 6.7% (vol/vol) ethanol for 4 wk. A cDNA tag that was overexpressed in the pancreas of rats fed ethanol was isolated. A 723-bp cDNA was cloned from a rat pancreatic cDNA library, which encodes a novel rat mitochondrial ATP synthase subunit 9, isoform 3 (ATP5G3), which is homologous to a human ATP5G3 gene. Real-time PCR demonstrated that all three nuclear gene isoforms (ATP5G1, ATP5G2, and ATP5G3) were consistently upregulated in the pancreas of alcohol-consuming rats, parallel with mitochondrial injury. The cellular response to mitochondrial damage and metabolic stress may reflect an adaptive process for mitochondrial repair in pancreatic acinar cells during chronic ethanol ingestion.

Role of γ-Subunit N- and C-Termini in Assembly of the Mitochondrial ATP Synthase in Yeast

2008 ◽  
Vol 377 (5) ◽  
pp. 1314-1323 ◽  
Author(s):  
Elke A. Dian ◽  
Panagiotis Papatheodorou ◽  
Kerstin Emmrich ◽  
Olga Randel ◽  
Andreas Geissler ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Γ Subunit ◽  
Mitochondrial Atp Synthase

scholarly journals TMPRSS4-dependent activation of the epithelial sodium channel requires cleavage of the γ-subunit distal to the furin cleavage site

2012 ◽  
Vol 302 (1) ◽  
pp. F1-F8 ◽  
Author(s):  
Christopher J. Passero ◽  
Gunhild M. Mueller ◽  
Michael M. Myerburg ◽  
Marcelo D. Carattino ◽  
Rebecca P. Hughey ◽  
...  
Keyword(s):  
Sodium Channel ◽  
Cleavage Site ◽  
Epithelial Sodium Channel ◽  
Cleavage Sites ◽  
Furin Cleavage ◽  
Channel Activation ◽  
Α Subunit ◽  
Γ Subunit ◽  
Furin Cleavage Site ◽  
Unique Mechanism

The epithelial sodium channel (ENaC) is activated by a unique mechanism, whereby inhibitory tracts are released by proteolytic cleavage within the extracellular loops of two of its three homologous subunits. While cleavage by furin within the biosynthetic pathway releases one inhibitory tract from the α-subunit and moderately activates the channel, full activation through release of a second inhibitory tract from the γ-subunit requires cleavage once by furin and then at a distal site by a second protease, such as prostasin, plasmin, or elastase. We now report that coexpression of mouse transmembrane protease serine 4 (TMPRSS4) with mouse ENaC in Xenopus oocytes was associated with a two- to threefold increase in channel activity and production of a unique ∼70-kDa carboxyl-terminal fragment of the γ-subunit, similar to the ∼70-kDa γ-subunit fragment that we previously observed with prostasin-dependent channel activation. TMPRSS4-dependent channel activation and production of the ∼70-kDa fragment were partially blocked by mutation of the prostasin-dependent cleavage site (γRKRK186QQQQ). Complete inhibition of TMPRSS4-dependent activation of ENaC and γ-subunit cleavage was observed when three basic residues between the furin and prostasin cleavage sites were mutated (γK173Q, γK175Q, and γR177Q), in addition to γRKRK186QQQQ. Mutation of the four basic residues associated with the furin cleavage site (γRKRR143QQQQ) also prevented TMPRSS4-dependent channel activation. We conclude that TMPRSS4 primarily activates ENaC by cleaving basic residues within the tract γK173-K186 distal to the furin cleavage site, thereby releasing a previously defined key inhibitory tract encompassing γR158-F168 from the γ-subunit.

scholarly journals Chloroplast Ribosomes Interact With the Insertase Alb3 in the Thylakoid Membrane

2021 ◽  
Vol 12 ◽  
Author(s):  
Bernd Ackermann ◽  
Beatrix Dünschede ◽  
Björn Pietzenuk ◽  
Bo Højen Justesen ◽  
Ute Krämer ◽  
...  
Keyword(s):  
Thylakoid Membrane ◽  
Molecular Mechanisms ◽  
Phylogenetic Analyses ◽  
Signal Recognition Particle ◽  
Ribosomal Subunit ◽  
Terminal Region ◽  
Hydrophobic Core ◽  
C Terminus ◽  
Terrestrial Life ◽  
Nascent Chain

Members of the Oxa1/YidC/Alb3 protein family are involved in the insertion, folding, and assembly of membrane proteins in mitochondria, bacteria, and chloroplasts. The thylakoid membrane protein Alb3 mediates the chloroplast signal recognition particle (cpSRP)-dependent posttranslational insertion of nuclear-encoded light harvesting chlorophyll a/b-binding proteins and participates in the biogenesis of plastid-encoded subunits of the photosynthetic complexes. These subunits are cotranslationally inserted into the thylakoid membrane, yet very little is known about the molecular mechanisms underlying docking of the ribosome-nascent chain complexes to the chloroplast SecY/Alb3 insertion machinery. Here, we show that nanodisc-embedded Alb3 interacts with ribosomes, while the homolog Alb4, also located in the thylakoid membrane, shows no ribosome binding. Alb3 contacts the ribosome with its C-terminal region and at least one additional binding site within its hydrophobic core region. Within the C-terminal region, two conserved motifs (motifs III and IV) are cooperatively required to enable the ribosome contact. Furthermore, our data suggest that the negatively charged C-terminus of the ribosomal subunit uL4c is involved in Alb3 binding. Phylogenetic analyses of uL4 demonstrate that this region newly evolved in the green lineage during the transition from aquatic to terrestrial life.

Identification of synapsin I peptides that insert into lipid membranes

2001 ◽  
Vol 354 (1) ◽  
pp. 57-66 ◽  
Author(s):  
James J. CHEETHAM ◽  
Sabine HILFIKER ◽  
Fabio BENFENATI ◽  
Thomas WEBER ◽  
Paul GREENGARD ◽  
...  
Keyword(s):  
Synaptic Vesicle ◽  
Synaptic Vesicles ◽  
Molecular Mechanisms ◽  
Phospholipid Bilayer ◽  
Synapsin I ◽  
Affinity Binding ◽  
Hydrophobic Core ◽  
Vesicle Membrane ◽  
High Affinity Binding ◽  
High Affinity

The synapsins constitute a family of synaptic vesicle-associated phosphoproteins essential for regulating neurotransmitter release and synaptogenesis. The molecular mechanisms underlying the selective targeting of synapsin I to synaptic vesicles are thought to involve specific protein–protein interactions, while the high-affinity binding to the synaptic vesicle membrane may involve both protein–protein and protein–lipid interactions. The highly hydrophobic N-terminal region of the protein has been shown to bind with high affinity to the acidic phospholipids phosphatidylserine and phosphatidylinositol and to penetrate the hydrophobic core of the lipid bilayer. To precisely identify the domains of synapsin I which mediate the interaction with lipids, synapsin I was bound to liposomes containing the membrane-directed carbene-generating reagent 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine and subjected to photolysis. Isolation and N-terminal amino acid sequencing of 125I-labelled synapsin I peptides derived from CNBr cleavage indicated that three distinct regions in the highly conserved domain C of synapsin I insert into the hydrophobic core of the phospholipid bilayer. The boundaries of the regions encompass residues 166–192, 233–258 and 278–327 of bovine synapsin I. These regions are surface-exposed in the crystal structure of domain C of bovine synapsin I and are evolutionarily conserved among isoforms across species. The present data offer a molecular explanation for the high-affinity binding of synapsin I to phospholipid bilayers and synaptic vesicles.

scholarly journals Structural model of F 1 –ATPase and the implications for rotary catalysis

2000 ◽  
Vol 355 (1396) ◽  
pp. 465-471 ◽  
Author(s):  
A. G. W. Leslie ◽  
J. E. Walker
Keyword(s):  
Crystal Structure ◽  
Conformational Changes ◽  
Structural Model ◽  
Change Mechanism ◽  
Γ Subunit ◽  
Catalytic Sites ◽  
Mechanism Of Catalysis ◽  
Rotary Catalysis ◽  
High Degree ◽  
Binding Change Mechanism

The crystal structure of bovine mitochondrial F 1 –ATPase is described. Several features of the structure are consistent with the binding change mechanism of catalysis, in which binding of substrates induces conformational changes that result in a high degree of cooperativity between the three catalytic sites. Furthermore, the structure also suggests that catalysis is accompanied by a physical rotation of the centrally placed γ–subunit relative to the approximately spherical α 3 β 3 sub–assembly.

Proteomics based detection of differentially expressed proteins in human osteoblasts subjected to mechanical stress

2013 ◽  
Vol 91 (2) ◽  
pp. 109-115 ◽  
Author(s):  
Fei-Fei Li ◽  
Fu-Lin Chen ◽  
Huan Wang ◽  
Shi-Bin Yu ◽  
Ji-Hong Cui ◽  
...  
Keyword(s):  
Mechanical Stress ◽  
Molecular Mechanisms ◽  
Bone Development ◽  
Expression Profiles ◽  
Differentially Expressed ◽  
Initiation Factor ◽  
Mechanical Stimuli ◽  
Mitochondrial Atp Synthase ◽  
Initiation Factor 2 ◽  
Protein Expression Profiles

Mechanical stress is essential for bone development. Mechanical stimuli are transduced to biochemical signals that regulate proliferation, differentiation, and cytoskeletal reorganization in osteoblasts. In this study, we used proteomics to evaluate differences in the protein expression profiles of untreated Saos-2 osteoblast cells and Saos-2 cells subjected to mechanical stress loading. Using 2-D electrophoresis, MALDI–TOF mass spectroscopy, and bioinformatics, we identified a total of 26 proteins differentially expressed in stress loaded cells compared with control cells. Stress loaded Saos-2 cells exhibited significant upregulation of 17 proteins and significant downregulation of 9 proteins compared with control cells. Proteins that were most significantly upregulated in mechanically loaded cells included those regulating osteogenesis, energy metabolism, and the stress response, such as eukaryotic initiation factor 2 (12-fold), mitochondrial ATP synthase (8-fold), and peptidylprolyl isomerase A (cyclophilin A)-like 3 (6.5-fold). Among the proteins that were significantly downregulated were those involved in specific signaling pathways and cell proliferation, such as protein phosphatase regulatory (inhibitor) subunit 12B (13.8-fold), l-lactate dehydrogenase B (9.4-fold), Chain B proteasome activator Reg (Alpha) PA28 (7.7-fold), and ubiquitin carboxyl-terminal esterase L1 (6.9-fold). Our results provide a platform to understand the molecular mechanisms underlying mechanotransduction.

scholarly journals Structure and conformational states of the bovine mitochondrial ATP synthase by cryo-EM

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Anna Zhou ◽  
Alexis Rohou ◽  
Daniel G Schep ◽  
John V Bason ◽  
Martin G Montgomery ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Catalytic Mechanism ◽  
Proton Translocation ◽  
Atp Synthesis ◽  
Bioinformatic Analysis ◽  
Mitochondrial Atp Synthase ◽  
A Subunit ◽  
Classification Of Images ◽  
Atp Synthases

Adenosine triphosphate (ATP), the chemical energy currency of biology, is synthesized in eukaryotic cells primarily by the mitochondrial ATP synthase. ATP synthases operate by a rotary catalytic mechanism where proton translocation through the membrane-inserted FO region is coupled to ATP synthesis in the catalytic F1 region via rotation of a central rotor subcomplex. We report here single particle electron cryomicroscopy (cryo-EM) analysis of the bovine mitochondrial ATP synthase. Combining cryo-EM data with bioinformatic analysis allowed us to determine the fold of the a subunit, suggesting a proton translocation path through the FO region that involves both the a and b subunits. 3D classification of images revealed seven distinct states of the enzyme that show different modes of bending and twisting in the intact ATP synthase. Rotational fluctuations of the c8-ring within the FO region support a Brownian ratchet mechanism for proton-translocation-driven rotation in ATP synthases.

scholarly journals Ocean acidification modulates expression of genes and physiological performance of a marine diatom

2015 ◽  
Vol 12 (18) ◽  
pp. 15809-15833 ◽  
Author(s):  
Y. Li ◽  
S. Zhuang ◽  
Y. Wu ◽  
H. Ren ◽  
F. Cheng ◽  
...  
Keyword(s):  
Ocean Acidification ◽  
Molecular Mechanisms ◽  
Carbon Fixation ◽  
Atp Synthesis ◽  
High Light Intensity ◽  
Marine Diatom ◽  
Physiological Performance ◽  
Light Regimes ◽  
Expression Of Genes ◽  
Mitochondrial Atp Synthase

Abstract. Ocean Acidification (OA) is known to affect various aspects of the physiological performance of diatoms, but there is little information on the underlining molecular mechanisms involved. Here, we show that in the model diatom Phaeodactylum tricornutum expression of the genes related to light harvesting, carbon acquisition and carboxylation, nitrite assimilation and ATP synthesis are modulated by OA. Growth and photosynthetic carbon fixation were enhanced by elevated CO2 (1000 μatm) under both constant indoor and fluctuating outdoor light regimes. The genetic expression of nitrite reductase (NiR) was up-regulated by OA regardless of light levels and/or regimes. The transcriptional expression of fucoxanthin chlorophyll a/c protein (lhcf type (FCP)) and mitochondrial ATP synthase (mtATP synthase) genes were also enhanced by OA, but only under high light intensity. OA treatment decreased the expression of β-carbonic anhydrase (β-CA) along with down-regulation of CO2 concentrating mechanisms (CCMs). Additionally, the genes for these proteins (NiR, FCP, mtATP synthase, β-CA) showed diel expressions either under constant indoor light or fluctuating sunlight. Thus, OA enhanced photosynthetic and growth rates by stimulating nitrogen assimilation and indirectly by down-regulating the energy-costly inorganic carbon acquisition process.

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