Cell Bio Exam 1
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How long are proteins are proteins typically?
How long is a cell typically?
The cell looks like _______ on the inside
What are cells made of? (4)
- 1. Proteins
- 2. Lipids
- 3. Nucleic Acids
- 4. small molecules
In the double layer membrane, the are the lipid tails attracted to each other?
No; hydrophobic tail is just repelled by water
What happens in the nucleolus?
The Golgi Apparatus is responsible for ______
- protein processing/ sorting
What does the smooth ER do?
produce and modify hydrophic molecules
What does the rough ER do?
ribosomes produce proteins that are soluble inside rough ER
site of aerobic ATP production
digestive; site of acidic hydrolases; breakdown of old cell parts and endo/phagocytosed materials
perform oxidative breakdown of fatty acids and other hydrophobic molecules
4 Pillars of Cell Biology
- 1. Biochem
- 2. Genetics
- 3. Microscopy
- 4. Molecular Biology
The study of structure, function, and interactions of biological macromolecules
- using electromagnetic radiation and lenses to visualize tiny objects
- -Limit of resolution is related to wavelength of light
Bright field microscopy
works on live cells, but no gives information about contents
makes use of fluorophores (absorb a high energy photon and emit a lower energy photon)
Immunofluorescence (3 steps)
- attach a fluorophore to a secondary antibody (ex GFP)
- 1. Fix cell (kill)
- 2. attach primary antibody
- 3. attach secondary antibody to primary
- -high resolution
- - inherently low contrast due to light makeup of cells
- - Solution: fox cells and stain using heavy atoms (eg. uranylacetate)
study of genes; relationship between genes and phenotypes
identification of genes that are responsible for known phenotypes
identification of phenotypes that are associated with known genes
use of recombinant DNA technology to manipulate and study genes and phenotypes
E. coli restriction 1- first restriction enzyme isolated. cuts DNA sequences in very precise predetermined locations
- -spin mixture at high G force for a few minutes
- -separate pellet from supernatant
- Gel electrophoresis
- uses charges to pull DNA molecules apart. The smaller the molecule, the faster it will move in a given time and the further it will get down the gel. Based on Molecular Weight not radius
Western Blot (steps)
- 1. gel elctrophoresis
- 2. transfer to introcellulose paper
- 3. add primary antibody to find molecule of interest
- 4. add secondary antibody to bind to primary. Chemi-lumocesence enzyme on antibody reacts chemically to produce a photon of light
primary protein structure
amino acid sequence
secondary protein structure
alpha helix, beta sheet; primary structure folding into a or b
tertiary protein structure
secondary elements come together to form a complex 3D strucure that is generally a functional protein
quaternary protein structure
complexes of proteins (eg. histones)
separation of molecules based on size, hydrophobicity, charge, or specific biological interactions
Ion exchange chromatography
Separates proteins based on charge. Beads coated with DEAE (positive) are placed in a tube. the solution of proteins goes through the tubes, and the negatively charged proteins attach to the beads and the cation proteins are eluted. a salt solution is used to elute the anions on the beads.
Gel Filtration Chromatography
- separates proteins based on size (Stokes Radius not molecular weight)
- -smaller proteins get stuck for a little bit in the depressions in the beads, so they elute more slowly than the larger proteins which easily go around the beads
- exploits biochemical interactions
- -molecules that bind to the protein of interest are covalently attached to the beads. The proteins not of interest are eluted first, then a salt change is used to elute the proteins of interest from the beads.
Actin monomers have ____ interactions with each other
actin and microtubles use _____ to produce work for movemetn
actin has a _____ end which is __ and a ______ end which is __
actin grows faster at the ______ end
barbed (+), Pointed (-)
actin is only an ATPase when ___________
it is in a filament (polymerized)
net addition at one end but net subtraction at the other
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