The plasma membrane, a protein-lipid bi-layer, is the primary barrier of a plant cell. It acts as a gatekeeper, regulating the flow of molecules and ions to maintain the cell's internal stability. This membrane facilitates nutrient uptake, waste removal, and maintains the cell's internal pressure, known as turgor pressure. It also detects signals from the environment, other cells, and pathogens.
Passive Transport
Definition: The spontaneous movement of molecules "downhill," from an area of higher free energy to an area of lower free energy or down a chemical potential gradient is called passive transport.
Driving Force: Driven by a concentration gradient (the difference in concentration between two areas). The process continues until equilibrium is reached, at which point there is no net movement of substances.
Follows Fick’s first law, where diffusion naturally occurs without the input of external energy.
Active Transport
Definition: The movement of molecules "uphill," against a gradient of chemical potential is called active transport.
Driving Force: This process is not spontaneous and requires an input of cellular energy to perform work. Often, this energy comes from the breakdown (hydrolysis) of ATP.
What is Chemical Potential?
Definition: Chemical potential is the partial molar Gibbs free energy of a substance. In simple terms, it's the energy change that occurs when one mole of a substance is added to a system, while keeping temperature and pressure constant.
Driving Force: It represents the "escaping tendency" of a substance. Molecules will naturally move from an area of higher chemical potential to an area of lower chemical potential.
Role in Transport: This energy difference, or gradient, is the driving force behind key processes like diffusion and osmosis, moving solutes across membranes until equilibrium is reached.
Key Factors
The chemical potential of a substance depends on several factors, including:
Concentration: As the concentration of a substance increases, its chemical potential also increases.
Temperature and Pressure: These physical conditions also influence the energy state of the molecules.
Electrical Potential: For charged molecules (ions), an electrical field also contributes to the chemical potential, creating an electrochemical potential.
Formula for Chemical Potential
For an ideal, dilute solution, the chemical potential (μ) is expressed by the equation:
Calculating the Driving Force
The direction and force of passive transport are determined by the difference in chemical potential (Δμ) between two areas.
For uncharged solutes (like sucrose): The driving force is simply the ratio of the solute's concentration on the inside and outside of the cell. If the concentration is higher outside, the solute will spontaneously move inward.
For charged solutes (ions, like K+): The driving force is influenced by both the concentration gradient and the electrical potential difference across the membrane. An ion can even be pushed against its concentration gradient if the electrical field is strong enough.
Diffusion
Transport of ions Across Membrane
Permeability: This is the extent to which a membrane allows a substance to pass through. It depends on both the membrane's composition and the chemical properties of the solute.
Effect on Diffusion: Membranes generally hinder diffusion, slowing down the process of reaching equilibrium. However, the membrane's permeability itself does not change the final equilibrium state, which is reached when the difference in electrochemical potential (Δμ) is zero.
Diffusion Potentials
Unequal Permeability: When different ions (like K+ and Cl-) diffuse across a membrane, they typically do so at different rates because the membrane's permeability to each ion is different.
Charge Separation: This difference in diffusion rates causes a slight separation of positive and negative charges, which instantly creates a diffusion potential (a voltage) across the membrane.
Neutrality: Although a potential exists, the actual number of unbalanced ions is chemically negligible. For example, a -100mV potential in a plant cell is created by an imbalance of just 1 extra negative ion for every 100,000.
The Nernst Equation
Predicting Equilibrium: The Nernst equation is a powerful tool used to determine if an ion is at equilibrium across a membrane.
The Balance: It states that at equilibrium, the electrical potential difference across the membrane exactly balances the concentration gradient of a specific ion.
Calculation: For diffusion of a specific ion:
Where:
R is the gas constant
T is the absolute temperature
z is the charge of the ion
F is Faraday’s constant
[ion]outside and [ion]inside are the concentrations of the ion outside and inside the cell, respectively
In typical biological settings at body temperature, this equation simplifies to:
This value represents the membrane potential at which the electrochemical forces acting on the ion are balanced, creating a state of equilibrium with no net movement.
Active vs. Passive Transport: A Nernst Test
The Nernst equation helps us distinguish between passive and active transport.
If an ion's actual measured internal concentration matches the concentration predicted by the Nernst equation, its transport is considered passive (driven by electrochemical potential).
If the measured concentration is significantly different from the predicted value, it indicates active transport is at play, requiring energy to move the ion against its electrochemical gradient.
The Role of Membrane Potential
All living cells have a membrane potential due to the unequal distribution of ions.
Plant cells often have a membrane potential much more negative than what can be explained by ion diffusion alone.
This extra voltage is generated by electrogenic pumps, which are active transport mechanisms that move a net electrical charge across the membrane.
In plant cells, the primary electrogenic pump is a H+-ATPase that actively pumps protons (H+) out of the cell. This process requires ATP and is a major determinant of the cell's membrane potential.
Impact: The voltage created by this pump in turn influences the passive diffusion of other ions, such as creating a strong electrical driving force for K+ to enter the cell.
How Membranes Transport Substances
Unlike simple artificial membranes, biological membranes are highly selective and much more permeable to ions, water, and large polar molecules. This is because they contain specialized transport proteins that facilitate the movement of specific substances across the membrane. These proteins fall into three main categories: channels, carriers, and pumps.
Channels (Enhanced diffusion)
Function: Channels are transmembrane proteins that act as selective pores. Substances diffuse through these pores very rapidly, with millions of ions passing through per second.
Transport Type: Transport through channels is always passive, meaning it follows the electrochemical gradient without needing extra energy.
Specificity: Their selectivity is based on the pore size and the charge of their inner lining.
Regulation: Channels are not always open. They have "gates" that open or close in response to various signals, such as changes in voltage (voltage-gated channels), chemical signals like hormones, or even mechanical force.
Directionality: Some channels are inwardly rectifying, allowing substances like K+ to move into the cell, while others are outwardly rectifying, allowing them to exit.
Carriers
Function: Carrier proteins do not form a continuous pore. They bind a specific substance, undergo a conformational change to move it across the membrane, and then release it on the other side.
Transport Rate: This process is much slower than channel transport, typically moving only a few hundred to a thousand molecules per second.
Specificity: Carriers are highly specific due to the need for a substance to bind to a specific site. They often transport specific ions, sugars, or amino acids.
Transport Type: Carrier-mediated transport can be either passive (also called facilitated diffusion) or active (Secondary active transport), where it's coupled with another energy-releasing event.
Categories: Carriers can be uniporters (passively transport a single substance) or co-transporters (actively transport two substances in a coupled fashion).
Membrane transport proteins
Pumps
Function: Pumps are membrane proteins that perform primary active transport, moving substances against their electrochemical gradient. This requires a direct input of energy, often from the breakdown of ATP or from light.
Energy Source: Directly uses energy from:
ATP Hydrolysis (e.g. H+- ATPase)
Oxidation-Reduction Reactions
Absorption of Light
Categorization: Pumps can be of two types:
- Electrogenic: moving a net electrical charge across the membrane (e.g., the H+-ATPase pumps H+ out, generating a voltage)
Electroneutral: no net charge movement (e.g., an H+/K+ pump exchanging H+ out for K+ in)
Significance: In plants, H+-ATPases are the most common electrogenic pumps. They actively pump protons out of the cell, which is a major factor in establishing the membrane potential. This creates Proton Motive Force (PMF), which is stored free energy used to drive secondary active transport.
Secondary Active Transport (Co-transport)
How it Works: This process uses the stored energy from one substance's downhill movement to drive the uphill transport of another. It's an indirect form of active transport.
The Driving Force: In plants, the strong proton gradient (or proton motive force) created by the H+-ATPases is the main source of energy for secondary active transport.
Types:
Symport: Two substances are moved in the same direction at the same time. Typically drives substrate uptake into the cytosol. For example, a proton and a sucrose molecule enter the cell together.
Antiport: Two substances are moved in opposite directions. Typically drives substrate export out of the cytosol. For instance, a proton moves into the cell while a sodium ion moves out.
Transport Kinetics
Studying Transport: Transport mechanisms can be analyzed using enzyme kinetics, as transport involves binding and dissociation at active sites. This helps us understand how transporters work and how they're regulated.
Key Parameters:
Vmax (Maximum Rate): This represents the maximum transport rate when all the transporter binding sites are occupied.
Vmax is approached when the substrate-binding site on the carrier is always occupied or when flux through the channel is maximal. The concentration of transporter, not the concentration of solute, becomes rate-limiting. Therefore, it indicates the number of transport proteins in the membrane.
Km (Concentration at Half Vmax): This represents nature of the binding sites. A low Km value means the transporter has a high affinity for its solute, which is characteristic of a carrier system.
Complex Systems: Many cells have multiple transport mechanisms for the same substance, such as both high- and low-affinity transporters, leading to complex kinetic curves. This often involves a mix of carrier-mediated (saturable) and simple diffusion (linear) processes.
General Principles of Membrane Transport
Proton Motive Force: In plants, most transport across membranes is driven by a single primary active transport system: H+-ATPases. These pumps use energy from ATP to actively transport protons (H+) out of the cell, creating an electrochemical potential gradient (the proton motive force).
Secondary Transport: This proton gradient is then used by a variety of secondary active transport proteins (symporters and antiporters) to move other ions and neutral substances.
Proton Cycling: Protons essentially "cycle" across the membrane: they are pumped out by primary pumps and flow back in via secondary transporters, driving the movement of other solutes.