Atp-dependent Translocation
Atp-dependent Translocation - Related Videos
01:31
ATP Yield
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2019
Cellular respiration produces 30 - 32 ATP per glucose molecule. Although most of the ATP results from oxidative phosphorylation and the electron transport chain (ETC), 4 ATP are gained beforehand (2 from glycolysis and 2 from the citric acid cycle). The ETC is embedded in the inner mitochondrial membrane and is comprised of four main protein complexes and an ATP synthase. NADH and FADH2 pass electrons to these complexes, which pump protons into the intermembrane space. This distribution of...
10:14
Biochemical Assays for Analyzing Activities of ATP-dependent Chromatin Remodeling Enzymes
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Cited by 4 •
2014
Here we describe biochemical assays that can be used to characterize ATP-dependent chromatin remodeling enzymes for their abilities to 1) catalyze ATP-dependent nucleosome sliding, 2) engage with nucleosome substrates, and 3) hydrolyze ATP in a nucleosome- or DNA-dependent manner.
08:32
The ATP Bioluminescence Assay
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2023
In fireflies, the luciferase enzyme converts a compound called luciferin into oxyluciferin, and produces light or “luminescence” as a result. This reaction requires energy derived from ATP in order to proceed, so researchers have exploited the luciferase-luciferin interaction to gauge ATP levels in cells. Given ATP’s role as the cell’s currency of energy, the ATP bioluminescence assay can provide insight into cellular metabolism and overall cell health.In this video, JoVE discusses cellular...
01:08
Hydrolysis of ATP
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2019
The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids. If one phosphate group is removed, a molecule of ADP—adenosine diphosphate—remains, along with inorganic phosphate. ADP can be further hydrolyzed to AMP—adenosine monophosphate—by the removal of a second...
12:27
Measuring Peptide Translocation into Large Unilamellar Vesicles
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Cited by 9 •
2012
This protocol details a method for the quantitative measure of peptide translocation into large unilamellar lipid vesicles. This method also provides information about the rate of membrane translocation and can be used to identify peptides that efficiently and spontaneously cross lipid bilayers.
FAQs
Each ATPase cycle changes the conformation of the ATPase or larger translocation complex. ATP binding, hydrolysis, and release of the resulting products occur as linked steps rather than isolated chemical events. These changes alter how the complex interacts with its cargo and transport route, allowing successive cycles to produce directional movement through a membrane or cellular channel.
Cargo binding connects the molecular substrate to the energy-converting translocation machinery. An ATPase or translocation complex must engage the protein, nucleic acid, or other cargo so that conformational changes generated during ATP turnover can be transmitted to it. This coupling helps determine what material enters the pathway and supports controlled movement rather than undirected displacement.
Directional movement allows cargo to reach a defined cellular destination instead of merely associating with a membrane or channel. Repeated conformational changes, driven by ATP turnover, create a coordinated progression through the transport route. This principle is important for maintaining compartmental organization, because proteins and other macromolecules can be delivered to particular cellular locations.
Its defining mechanistic feature is the direct use of ATP hydrolysis to drive conformational changes in a motor or translocation complex. The energy input is therefore coupled to cargo movement through a membrane or channel. This distinguishes the process from movement that would not rely on repeated ATP binding, hydrolysis, and product release by the transport machinery.
Reconstitution allows researchers to examine a transport pathway as a defined molecular system and relate ATP turnover to cargo movement. It can support analysis of the responsible ATPase or translocation complex, its cargo interactions, and the conformational changes associated with transport. Such studies help characterize how a pathway operates without relying only on its broader cellular role.
The mechanism supports several major transport contexts, including protein import into organelles, passage through secretion systems, and insertion of proteins into membranes. It also contributes to remodeling or transport of macromolecular complexes. These applications show that the same energy-coupling principle can handle different cargo types and operate across distinct cellular transport pathways.
Studies can reveal how molecular motors convert ATP turnover into mechanical or positional changes, how translocation complexes engage cargo, and how transport pathways are organized. Researchers can use these observations to characterize motors and reconstitute pathways in controlled systems. The resulting mechanistic information connects molecular activity with the maintenance of cellular compartments and transport routes.