Cell Bio Exam 2

The ___ and ___ of a cell is define by its protein expression

structure and function

What are the 3 steps to isolate proteins from cells or tissue

Step 1. Break open cells to release their contents (Lysate)
Step 2. Remove unwanted parts of the cell (Centrifuge)

Step 3. Purify specific protein ( Electrophoresis, Chromatography)

What are some methods to isolate proteins from the cell (Step 1)

• Ultrasounds, pressure shear force

• This produces Cell Lysate

___ and ___ are used to denature proteins and apply uniform negative charge prior to loading

Reagents and heat

Solve the problem

Define Western Blotting

Western Blotting is a lab technique used to detect a specific protein in a sample using antibodies

ELISA (Enzyme-Linked Immunosorbent Assay) is the most reliable way to ___ a specific protein

Quantify

What 2 pathways constitute a cells metabolism

Catabolic and Anabolic

Understand all terms in the free energy equation

ΔG = ΔG° + RT ln Q

ΔG = free energy change (spontaneity)

ΔG° = standard free energy change

R = 8.314 J/mol·K

T = temperature (K)

Q = [products]/[reactants]

ΔG < 0 → spontaneous

ΔG > 0 → non-spontaneous

ΔG = 0 → equilibrium

Reaction ___ can drive energetically unfavorable reactions

Coupling

What is the main purpose of an Enzyme

To reduce the energy needed to initiate spontaneous reactions

Enzymes convert ___ to ___ in a repeatable cycle

substrates to products

Understand the terms in the Michaelis-Menten Equation
v = (Vmax [S]) / (Km + [S])

v = (Vmax [S]) / (Km + [S])

v = reaction rate

Vmax = max rate (enzyme saturated)

[S] = substrate concentration

Km = [S] at 1/2 Vmax (affinity indicator)

Low Km = high affinity

High Km = low affinity

Gene expression is carried out through the ___

Central Dogma

Define Gene Expression

The process by which a gene makes a product that is useful to the cell by directing the synthesis of a protein or an RNA molecule with a characteristic activity