Perfect your technique
Testing necessity typically involves this kind of experiment
What is knocking something out, mutating something
When proteins were mutated to remove the NLS, they localized here
cytosol
This short N-terminal sequence, rich in hydrophobic amino acids, targets a protein to the ER.
What is the signal peptide (or signal sequence)?
This small GTPase initiates COPII vesicle formation at the ER membrane.
What is Sar1?
This is the ligand for the receptor tyrosine kinase receptor discussed in class
What is EGF (Epidermal Growth Factor)?
Differential centrifugation is used to fractionate/separate these structures from one another based on differences in size and density
What are organelles?
This protein hydrolyzes the GTP on Ran during nuclear transport.
What is Ran?
If SRP were missing during translation, secretory proteins would accumulate in this location.
What is the cytosol?
In this model of inter-Golgi transport, only enzymes but not cargo are transported in vesicles
What is cisternal maturation model?
These two types of molecular switches are commonly used to control signaling pathways.
What are GTP-binding proteins and protein kinases?
Column chromatography is commonly used to separate these molecules based on differences in properties like size, charge, or affinity.
What are proteins?
This protein stimulates the dissociation of GDP on Ran
What is Ran GEF?
Deleting the KDEL sequence from a soluble ER resident protein causes it to localize here
What is the extracellular space?
These GTP-binding proteins are present on both the vesicle and the target membrane
What are Rab proteins?
In the MAP kinase cascade (RAF → MEK → ERK), if MEK is constitutively active in a cancer patient, this type of drug would most effectively inhibit tumor growth.
What is an ERK (MAP kinase) inhibitor?
This advancement in microscopy allows visualization of proteins at resolutions of up to 2 nm.
What is super-resolution microscopy?
If Ran were permanently locked in the GDP-bound form, this critical step at the nuclear periphery would fail during nuclear import
What is the release of cargo from importin?
This type of membrane protein has its N-terminus in the ER lumen and C-terminus in the cytosol and uses a cleaved N-terminal signal sequence.
What is a Type I membrane protein?
In the Rothman in vitro fusion assay, addition of a non-hydrolyzable ATP analog would most directly block this step of fusion.
What is Disassembly of v-SNARES and t-SNARES from each other?
An adaptor protein binds the phosphorylated receptor and brings this type of protein to the membrane, which then activates Ras.
What is Ras GEF, aka SOS?
If three purified proteins can recreate a biological process in a test tube, those proteins are considered this.
What is sufficient?
If Ran lost its GTPase activity and stayed permanently in the GTP-bound form, importin would remain bound to this protein, preventing new rounds of cargo import.
What is Ran.GTP?
Integral membrane proteins that do not have transmembrane domains are anchored to the membrane via this modification.
What is a lipid anchor such as a GPI anchor?
If a vesicle contains cargo proteins but fails to dock at any membrane despite correct SNAREs being present, which one of these two types of protein is most likely defective?
What is a Rab GTPase or Rab effector?
MAP kinase is constitutively active. A Ras inhibitor has no effect. You don't know where the mutation lies.
Should you try inhibiting something upstream or downstream of Ras?
What is downstream of Ras?