BIOMOLECULES o Flagella – propel cell

Midterm Reviewer o Cell Envelope – varies with

type of bacteria
Lecture 1 - Prokaryotic vs. Eukaryotic Cell
Biochemistry Structures

I. Introduction to the Cell

Definition of Biochemistry
- Biology – study of life
- Chemistry – composition, properties
and transformations of matter
- Biochemistry – study of the
chemistry of life; biologically 2. Eukaryotic – contain true nuclei and
important elements and compounds are much larger and complex
internally
The Cell
- The basic unit of life in all forms of 6-Kingdom Classification
living organisms
- Types of Cell
1. Prokaryotic – simplest type of cells
Gram-negative Gram-positive
bacteria bacteria
- Outer membrane - No outer
- Peptidoglycan membrane
layer - Thicker
peptidoglycan
Cyanobacteria Archaebacteria
- Gram-negative - No outer
- Tough membrane
peptidoglycan - Peptidoglycan
- Photosynthetic outside plasma
pigments membrane Three Domains of Life
- Cell Structures

- Organisms in all three domains of

o Ribosomes – smaller than
eukaryotic ribosomes, for
protein synthesis
o Nucleoid – contains circular
DNA
o Pili – points of adhesion
life share this property: they are all
made up of cells
The Hierarchical Organization of The
Multicellular Organism
 Universal Features of Living Cells

Structural Hierarchy in the Molecular

Organization of the Cell

Subcellular Fractionation of Tissue

Source of Energy

Processes That Regulate Cell Contents

1. Diffusion – high to low solute
2. Osmosis – solvent into higher
- Humans are organotrophs – take solute (thru semipermeable
up organic compounds. membrane)
3. Dialysis – solute across
semipermeable membrane due to
concentration gradient
 4. Surface Tension – attraction of
liquid molecules (cohesion)
Lecture 1: Activity
1. What is the pH of a HCl (aq) solution
with an [H+] = 1.5x10^-4?
2. What is the pOH of the HCl (aq)
Tonicity
solution with an [H+] = 1.5x10^-4?
3. Calculate the pKa of lactic acid,
given that when the concentration of
lactic acid is 0.020M and the
concentration of lactate is 0.175M,
the pH is 4.80.
4. Calculate the pH of a mixture of
0.15M acetic acid and 0.25M sodium
acetate. The pKa of acetic acid is
4.76.
Sucrose Density Gradient Centrifugation
A. Centrifugation Types
5. Calculate the ratio of the
concentrations of acetate and acetic
acid required in a buffer system of
pH 4.30.
- Describes how an extracellular
solution can change the volume of a
cell by affecting osmosis
- Osmolarity – total solute
concentration of the solution.
o Low Osmolarity – greater
water molecules, lower solute
particles
o High Osmolarity – fewer
water molecules, higher
solute particles
- Water will move from side of lower
osmolarity to side of higher
osmolarity.
- Hypotonic (situation) – extracellular
fluid has lower osmolarity than the
fluid inside the cell > water enters
the cell – swells, may burst
- Hypertonic (situation) –
extracellular fluid having a higher
osmolarity than the cell’s cytoplasm
> water is released from the cell –
shrink, shriveled
- Isotonic (situation) – extracellular
fluid has the same osmolarity as the
cell > no net movement – retain
shape
- Two Centrifugation Methods:
1. Differential Centrifugation
o Simple form
o Performed at different
centrifugal speeds
 oSeparate particles with
different sedimentation
coefficients
2. Density Gradient Centrifugation
o More complicated
o Rate-zonal Centrifugation
o Isopycnic Centrifugation
*Differential Centrifugation
- Gradually increasing the centrifugal
speed to separate particles with
different sedimentation coefficients
- Disadvantage: uneven
sedimentation, needs to be
resuspended
*Rate-zonal Centrifugation
- Formation of bands, density of
gradients is lower than samples
- Control of centrifugal time is
important
- Separate particles of similar density
with different molecular weight (e.g.
proteins)
- Tip of the pipette should be at the
bottom of centrifugal tubes
- Gradient must be cold, used as soon
as possible
- Gradient Forming Instruments
2. Centrifugation
- Add the sample on top of the
gradient (sucrose)
- Centrifuge must be balance prior:
pre-experiment (centrifugal time,
force)
- Centrifugation > bands generation
3. Separation and Elution
- Two ways:
o Bottom-up – Gradient
fractionator
o Top-down – pipette
- Elusion – further purification,
separate sucrose from particles

*Isopycnic Centrifugation
- Separation based on density
- Used to separate particles with
similar molecular weight but different
densities (e.g. nucleic acids,
organelles)

B. Sucrose Density Gradient

Centrifugation
1. Gradient Preparation
- Sucrose solution (66% w/w) >
diluted with different concentrations
- Sucrose must be filtered and
analyzed with refractometer
- Lower concentration succeeded by
higher concentrations