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Membrane Transport Proteins: Membrane proteins whose primary function is to facilitate the transport of molecules across a biological membrane. Included in this broad category are proteins involved in active transport (Biological transport, Active), facilitated transport and Ion channels.

Primary Active Transport

JoVE 10706

In contrast to passive transport, active transport involves a substance being moved through membranes in a direction against its concentration or electrochemical gradient. There are two types of active transport: primary active transport and secondary active transport. Primary active transport utilizes chemical energy from ATP to drive protein pumps that are embedded in the cell membrane. With energy from ATP, the pumps transport ions against their electrochemical gradients—a direction they would not normally travel by diffusion. To understand the dynamics of active transport, it is important to first understand electrical and concentration gradients. A concentration gradient is a difference in the concentration of a substance across a membrane or space that drives movement from areas of high concentration to areas of low concentration. Similarly, an electrical gradient is the force resulting from the difference between electrochemical potentials on each side of the membrane that leads to the movement of ions across the membrane until the charges are similar on both sides of the membrane. An electrochemical gradient is created when the forces of a chemical concentration gradient and electrical charge gradient are combined. One important transporter responsible for maintaining the electrochemical gradient in cells is the sodium-potassium pump. The pr

 Core: Biology

Reconstitution of Membrane Proteins

JoVE 5693

Reconstitution is the process of returning an isolated biomolecule to its original form or function. This is particularly useful for studying membrane proteins, which enable important cellular functions and affect the behavior of nearby lipids. To study the function of purified membrane proteins in situ, they must be reconstituted by integrating them into an artificial lipid membrane.

 Biochemistry

Facilitated Transport

JoVE 10705

The chemical and physical properties of plasma membranes cause them to be selectively permeable. Since plasma membranes have both hydrophobic and hydrophilic regions, substances need to be able to transverse both regions. The hydrophobic area of membranes repel substances such as charged ions. Therefore, such substances need special membrane proteins to cross a membrane successfully. In the process of facilitated transport, also known as facilitated diffusion, molecules and ions travel across a membrane via two types of membrane transport proteins: channels and carrier proteins. These membrane transport proteins enable diffusion without requiring additional energy. Channel proteins form a hydrophilic pore through which charged molecules can pass through, thus avoiding the hydrophobic layer of the membrane. Channel proteins are specific for a given substance. For example, aquaporins are channel proteins that specifically facilitate the transport of water through the plasma membrane. Channel proteins are either always open or gated by some mechanism to control flow. Gated channels remain closed until a particular ion or substance binds to the channel, or some other mechanism occurs. Gated channels are found in the membranes of cells such as muscle cells and nerve cells. Muscle contractions occur when the relative concentrations of ions on the interior and

 Core: Biology

Short-distance Transport of Resources

JoVE 11097

Short-distance transport refers to transport that occurs over a distance of just 2-3 cells, crossing the plasma membrane in the process. Small uncharged molecules, such as oxygen, carbon dioxide, and water, can diffuse across the plasma membrane on their own. In contrast, ions and larger molecules require the assistance of transport proteins due to their charge or size. Transport across membranes also occurs within individual cells, playing a variety of essential roles for the plant as a whole. Resources are transported into and out of the central vacuole within each plant cell One of the roles of the large central vacuole of a plant cell is the storage of resources. Active and passive transport proteins are found in the vacuolar membrane, or tonoplast, just as they are found in the plasma membrane of the cell, and they regulate the movement of solutes between the cytoplasm and vacuole. Sugar can be stored for later, ions are sequestered from the cytoplasm, and protons, in particular, are pumped into the vacuole, creating an acidic environment for breaking down unwanted or toxic substances that enter the cell. Movement across the tonoplast controls turgor pressure In addition to its role in storage, the vacuole generates turgor pressure - a force that pushes the plasma membrane against the cell wall -

 Core: Biology

Electron Transport Chains

JoVE 10742

The final stage of cellular respiration is oxidative phosphorylation, which consists of (1) an electron transport chain and (2) chemiosmosis.

The electron transport chain is a set of proteins and other organic molecules found in the inner membrane of mitochondria in eukaryotic cells and the plasma membrane of prokaryotic cells. The electron transport chain has two primary functions: it produces a proton gradient—storing energy that can be used to create ATP during chemiosmosis—and generates electron carriers, such as NAD+ and FAD, that are used in glycolysis and the citric acid cycle. Generally, molecules of the electron transport chain are organized into four complexes (I-IV). The molecules pass electrons to one another through multiple redox reactions, moving electrons from higher to lower energy levels through the transport chain. These reactions release energy that the complexes use to pump H+ across the inner membrane (from the matrix into the intermembrane space). This forms a proton gradient across the inner membrane. NADH and FADH2 are reduced electron carriers produced during earlier cellular respiration phases. NADH can directly input electrons into complex I, which uses the released energy to pump protons into the intermembrane space. FADH2 inputs electrons into complex II, the only co

 Core: Biology

Secondary Active Transport

JoVE 10707

One example of how cells use the energy contained in electrochemical gradients is demonstrated by glucose transport into cells. The ion vital to this process is sodium (Na+), which is typically present in higher concentrations extracellularly than in the cytosol. Such a concentration difference is due, in part, to the action of an enzyme “pump” embedded in the cellular membrane that actively expels Na+ from a cell. Importantly, as this pump contributes to the high concentration of positively-charged Na+ outside a cell, it also helps to make this environment “more positive” than the intracellular region. As a result, both the chemical and electrical gradients of Na+ point towards the inside of a cell, and the electrochemical gradient is similarly directed inwards. Sodium-glucose cotransporters (SGLTs) exploit the energy stored in this electrochemical gradient. These proteins, primarily located in the membranes of intestinal or kidney cells, help in the absorption of glucose from the lumen of these organs into the bloodstream. In order to function, both an extracellular glucose molecule and two Na+ must bind to the SGLT. As Na+ migrates into a cell through the transporter, it travels with its electrochemical gradient, expelling energy that the protein uses to move glucose ins

 Core: Biology

Chemiosmosis

JoVE 10743

Oxidative phosphorylation is a highly efficient process that generates large amounts of adenosine triphosphate (ATP), the basic unit of energy that drives many processes in living cells. Oxidative phosphorylation involves two processes—electron transport and chemiosmosis. During electron transport, electrons are shuttled between large complexes on the inner mitochondrial membrane and protons (H+) are pumped across the membrane into the intermembrane space, creating an electrochemical gradient. In the next step, protons flow back down their gradient into the mitochondrial matrix via ATP synthase, a protein complex embedded within the inner membrane. This process, called chemiosmosis, uses the energy of the proton gradient to drive the synthesis of ATP from adenosine diphosphate (ADP). The electron transport chain is a series of complexes that transfer electrons from electron donors to electron acceptors via simultaneous reduction and oxidation reactions, otherwise known as redox reactions. At the end of the chain, electrons reduce molecular oxygen to produce water. The shuttling of electrons between complexes is coupled with proton transfer, whereby protons (H+ ions) travel from the mitochondrial matrix to the intermembrane space against their concentration gradient. Eventually, the high concentration of protons in the interm

 Core: Biology

The Apoplast and Symplast

JoVE 11106

Plant growth depends on its ability to take up water and dissolved minerals from the soil. The root system of every plant is equipped with the necessary tissues to facilitate the entry of water and solutes. The plant tissues involved in the transport of water and minerals have two major compartments - the apoplast and the symplast. The apoplast includes everything outside the plasma membrane of living cells and consists of cell walls, extracellular spaces, xylem, phloem, and tracheids. The symplast, in contrast, consists of the entire cytosol of all living plant cells and the plasmodesmata - which are the cytoplasmic channels interconnecting the cells. There are several potential pathways for molecules to move through the plant tissues: The apoplastic, symplastic, or transmembrane pathways. The apoplastic pathway involves the movement of water and dissolved minerals along cell walls and extracellular spaces. In the symplastic route, water and solutes move along the cytosol. Once in this pathway, materials need to cross the plasma membrane when moving from cell to neighboring cell, and they do this via the plasmodesmata. Alternatively, in the transmembrane route, the dissolved minerals and water move from cell to cell by crossing the cell wall to exit one cell and enter the next. These three pathways are not mutually exclusive, and some solutes may use more than on

 Core: Biology

What Are Osmoregulation and Excretion?

JoVE 11001

Organisms must keep bodily fluids at a constant temperature and pH while maintaining specific solute concentrations in order to support life functions. Osmoregulation is the process that balances solute and water levels.

Osmosis is the tendency of water to move from solutions with lower ion concentrations, or osmolarities, to those with higher ion concentrations. Osmosis occurs in response to differences in the molecular concentrations of solutions separated by a semipermeable membrane. Bodily fluids, which are separated by such membranes, contain water, non-electrolytes, and electrolytes—solutes that dissolve into ions in water. Both electrolytes and non-electrolytes influence osmotic balance. However, since the more important factor to osmosis is solute number, rather than size, the contribution of electrolytes is more significant. Unlike water, electrolytes cannot diffuse passively through membranes but rely on facilitated diffusion and active transport. In facilitated diffusion, protein-based channels move solutes across membranes. Conversely, energy is used to move ions against concentration gradients in active transport. When animals ingest food, material that cannot be used is excreted from the body. Excretory systems in nature involve tradeoffs between conserving energy and water. Nitrogen is among the most signific

 Core: Biology

Protein Associations

JoVE 10704

The cell membrane—or plasma membrane—is an ever-changing landscape. It is described as a fluid mosaic as various macromolecules are embedded in the phospholipid bilayer. Among the macromolecules are proteins. The protein content varies across cell types. For example, mitochondrial inner membranes contain ~76%, while myelin contains ~18% protein content. Individual cells contain many types ofbrane proteins—red blood cells contain over 50—and different cell types harbor distinct membrane protein sets. Membrane proteins have wide-ranging functions. For example, they can be channels or carriers that transport substances, enzymes with metabolic roles, or receptors that bind to chemical messengers. Like membrane lipids, most membrane proteins contain hydrophilic (water-loving) and hydrophobic (water-fearing) regions. The hydrophilic areas are exposed to water-containing solution inside the cell, outside the cell, or both. The hydrophobic regions face the hydrophobic tails of phospholipids within the membrane bilayer. Membrane proteins can be classified by whether they are embedded (integral) or associated with the cell membrane (peripheral). Most integral proteins are transmembrane proteins, which traverse both phospholipid layers, spanning the entire membrane. Their hydrophilic regions extend from both sides of the membrane, facing cytosol on

 Core: Biology
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