Unit 4 Exploring Proteins
Unit 5 and Unit 6
Unit 7 Carbohydrates
Unit 8 Nucleic Acids
Unit 9 DNA Synthesis
100
Define Cell Lysis.
The breaking open of a cell's outer membrane/wall to release its internal contents.
100
The enzyme classification of Kinase
What is transferase?
100
What is the structure of glucose?
100
Complementary RNA Sequence:
5' - GCACGCTTAGCT - 3'
5' - AGCUAAGCGUGC - 3'
100
Direction of DNA synthesis
&
Type of directional replication that cells use and why
5' end to 3' end
Bidirectional replication because it's more efficient
200
What are the steps of 2D Electrophoresis? (Include what each detects)
First Dimension: pI is detected via Isoelectric Focusing
Second Dimension: Size is detected via SDS-PAGE
200
This model of Enzyme-Substrate Binding assumes that the enzyme must be flexible in order to fit the substrate.
The ‘Induced-Fit Model’.
200
Difference between glycosidases and glycosyl transferases
glycosidases: hydrolyzes glycosidic bonds
glycosyl transferases: condensation reaction forming glycosidic bonds
200
DNA's secondary structure of polynucleotide chains
What are antiparallel and noncovalently (Hydrogen) bonded?
200
What did the Meselson-Stahl experiment prove?
the semiconservative method was used in DNA replication
300
How is the pH gradient established in isoelectric focusing, and what stops a given protein's migration?
It is established through the electrophoresis of a mix of polyampholytes.
The protein migrates until it reaches a pH that is equal to it's own pI.
300
What kind of inhibitor would decrease Km and Vmax?
Uncompetitive Inhibitor
300
What is the difference between proteglycans and glycoproteins (what are their features, where are they synthesized, etc.)?
proteoglycan: core protein covalently attached to GAG, synthesis on the ER, secreted, surface bound
glycoprotein: protein is the major component determining features, shorter than GAGs, glycosylation contributes to activity, sorting
300
Sequence of sugar and phosphate
What is the backbone of nucleic acids?
300
Define exonuclease and endonuclease
Exonuclease: enzymes that remove nucleotides from the ends of DNA/RNA chain
Endonuclease: enzymes that remove nucleotides from anywhere in DNA/RNA chain
400
What is ELISA? Define the two types, what they detect, and what is attached to their wells.
Elisa is a test that detects and measures specific antibodies, antigens, proteins, etc. in fluid samples.
The two types are Indirect and Sandwich.
Indirect detects patient [serum] antibodies. Sandwich detects the antigen in the sample.
In Indirect ELISA the known antigen is attached to the well. In Sandwich ELISA, the capture antibody is attached, along with another antibody that is bound to another epitope on the captured antigen.
400
This refers to the protein moiety of an enzyme that requires a cofactor coenzyme and is functional
Holoenzyme
400
Oligosaccharide linkages and where they are synthesized?
O-linked --> Ser, Thr, synthesized in the golgi
N-linked --> Asn, synthesized in the ER and then the golgi
400
Difference between type I topoisomerases and type II isomerases
Type I isomerases: single stranded, intermediate nicks, release supercoils
Type II isomerases: double stranded intermediate nicks, release or introduce supercoils
400
What type of intrinsic endonuclease/exonuclease does DNA polymerase have?
3' to 5' exonuclease activity
500
Give the five steps of protein purification. Explain why the target protein yield reduces through this process.
Choose a rich source --> Enrich the target fraction --> Purify the protein --> Monitor and quantify each step --> Analyze structure and function
500
If a point mutation in a small GTPase like RAS completely prevents its interaction with GAPs, what is the expected impact on its function?
RAS will remain persistently in its active, GTP-bound conformation due to impaired GTP hydrolysis.
Without GAP interaction to stimulate intrinsic GTPase activity, GTP hydrolysis proceeds extremely slowly, keeping locked in its state.
500
What is a lectin and what are the different functions in different species?
Lectin: carbohydrate binding protein
Animals: cell-cell recognition, signaling, adhesion, removal
viral: binding to host
bacteria: adhesion to host
Plant: protection from microbes
500
Differences between DNA double helix and protein alpha helix
DNA: hydrogen bonds are perpendicular, flexible
Alpha - helix: hydrogen bonds are parallel, stiff
500
List things that DNA polymerase needs (3 items)
1) DNA template
2) free 3' OH end through a primer
3) activated bases (phosphate needed)