BIOL 360: Molecules Flashcards
What enzyme keeps phosphatidylserine positioned on the cytosolic face of the plasma membrane of a living cell?
Flippase.
What is sphingosine?
A molecule with a 3-C backbone, linked to a fatty chain on one end, a hydroxyl group at either end, and an amine group at the central C; forms the basis of sphingolipids and glycolipids.
What is a lamellipodium?
A 2-dimensional cell protrusion, created by a 2-D network of actin filaments spreading outward from the cell.
Which actin homologues in bacterial cells help to determine cell shape?
MreB and Mbl.
What is YFP (yellow fluorescent protein)?
A fluorescent protein used to transgenically label specific proteins for visualization by fluorescence microscopy (emits yellow light).
What is the G0 phase of the cell cycle?
A prolonged interphase in a cell that stays in G1 for a very long time (or forever).
How do the S4 helices in voltage-gated Na+ channels trigger the conformational change that opens the channel?
When the membrane depolarizes, the extracellular side of the membrane becomes negatively charged relative to the cytosolic side, so the positively charged amino acids on the S4 helices experience an electrostatic pulling force as they are attracted to the opposite side of the membrane.
What happens in S phase of the cell cycle?
Genetic material is replicated (S = synthesis of genetic material).
What is a triacylglycerol?
A class of esters derived from glycerol and 3 fatty acids, stored in lipid droplets where they arrange in monolayers; the main form in which fat is stored.
What is a low-density lipoprotein (LDL)?
A spherical particle containing about 1,500 cholesterol molecules esterified to fatty acid tails, used to tranport cholesterol in the blood.
What are Cdc6 and Cdt1?
Proteins that collaborate with the ORC in late M/early G1 to help load inactive DNA helicases onto the DNA next to the origin of replication.
What is the main structural difference between cytoplasmic and axonemal (ciliary) dyneins?
Cytoplasmic dyneins are homodimers; axonemal dyneins are heterodimers or heterotrimers.
What is ParM?
An actin homologue in bacterial cells that helps to segregate plasmids during plasmid replication.
What is p21?
A Cdk inhibitor protein (CKI) in mammalian cells that suppresses G1/S- and S-Cdk activity following DNA damage.
What is an APC/C?
An anaphase-promoting complex (cyclosome): a complex present in anaphase that catalyzes the destruction of M- and S-cyclins and securin.
What adaptor protein functions as a coincidence detector during clathrin coat assembly?
AP2.
Why is yeast an especially good model for studying genes involved in the cell cycle?
It can proliferate in a haploid state, so any mutant genes introduced will have their full effect (no backup wild-type gene to compensate).
What is the first interaction that occurs between a vesicle and its target membrane?
Binding between the vesicle’s Rab-GTP and the target membrane’s Rab effector.
What is a farnesyl anchor?
A lipid anchor that irreversibly attaches a protein to the cytosolic side of the plasma membrane with a thioether linkage between a prenyl group and a Cys residue in the protein.
What is a morphogen?
An extracellular signal that stimulates uncommitted stem cells to differentiate.
What is fimbrin?
A protein that packs actin filaments tightly together in tight parallel bundles, allowing no room for other proteins between the filaments.
What is Cas9?
An engineered Cas mutant that does not cleave double-stranded DNA but instead inhibits it (allowing researchers to turn off specific genes).
What is condensin?
A protein complex made up of 2 Smc molecules and 3 CAP proteins with a hinge domain and an ATPase domain, which forms a ring around DNA to help condense chromosomes during mitosis.
What is Cdh1?
A protein that maintains APC/C activity after anaphase and throughout G1 until inhibited by Cdk activity.
What happens to a nuclear export receptor once it reaches the cytosolic side of a nuclear pore complex?
Ran-GAP in the cytosol causes receptor-bound Ran-GTP to hydrolyze its GTP to GDP, which causes the receptor to release both Ran-GDP and its cargo protein (as Ran-GTP is no longer present to promote its binding to either).
What is an Arp2/3 complex?
A complex of two actin-related proteins (ARPs) that nucleates actin filament growth from the minus end to create branching actin filaments.
What is Cas? How does it work?
A protein involved in the bacterial CRISPR system:
- RNA transcribed from CRISPR locus binds a Cas protein
- New virus injects double-stranded DNA
- If complementary, RNA mediates binding between Cas and foreign DNA
- Cas cleaves foreign DNA
What is Cdc25 phosphatase?
A phosphatase that removes an inhibitory phosphate group (added by Wee1 kinase) from a Cdk-cyclin complex to activate it.
What is a GDI (Rab-GDP dissociation inhibitor)?
A protein that binds inactive Rab-GDP and keeps it soluble in the cytosol until Rab is activated.
Why is it important to use conditional mutants when studying genes involved in the cell cycle?
The cell cycle needs to function correctly to generate enough cells to work with, so conditional mutants allow researchers to turn genes on long enough to get a good culture, then off to study the effects of the mutations.
How are intermediate filaments assembled?
- 2 α-helical monomers form a parallel coiled-coil dimer
- 2 dimers form a staggered, antiparallel tetramer
- 8 tetramers associate laterally into bundles
- Bundles link end-to-end to form filaments
How long does S phase take in E. coli cells?
About 20 minutes.
What type of linkages are formed between membrane-bound proteins and myristoyl, palmitoyl, and farnesyl anchors?
- Myristoyl: amide (irreversible)
- Palmitoyl: thioester (reversible)
- Farnesyl: thioether (irreversible)
How do aquaporins prevent protons from travelling through their channels?
Protons usually travel by hopping between water molecules (briefly converting them to H3O+), but strategically placed Asn residues interact with H2O within the channel so that the O has no free electrons to participate in proton relay (and the proton has no choice but to bounce back to where it came from).
Which kinase is needed to fully activate a Cdk-cyclin complex?
CAK (Cdk-activating kinase).
True or false: All linkages between lipid anchors and their associated membrane-bound proteins are post-translational.
True.
What are the 3 main protrusive structures in cells used for whole-cell locomotion?
- Filopodia
- Lamellipodia
- Pseudopodia
What is treadmilling?
Movement of actin filaments by simultaneous polymerization at the plus end and depolymerization at the minus end.
How does the inactivation gate work in voltage-gated Na+ channels?
As soon as membrane depolarization triggers the opening of the channel, the inactivation gate automatically swings shut to keep the channel from transporting more Na+ even while the membrane is still depolarized, ensuring that membrane depolarization is propagated in one direction only.
What is p27?
A Cdk inhibitor protein (CKI) in mammalian cells that suppresses G1/S- and S-Cdk in G1 and helps cells withdraw from the cell cycle when they terminally differentiate.
How do lysosomes protect themselves from degradation by their own enzymes?
Lysosomal membrane proteins are very highly glycosylated on the inner face of the membrane, so they protect the membrane from damage.
What are kinesins?
A group of motor proteins that move along microtubules toward the plus end.
What are the 2 functional phases of mitosis?
Cdk activity (early) and APC/C activity (late).
What is unique about COPI coat assembly (vs. COPII & clathrin)?
The components of their inner and outer coat layers are recruited as a preassembled complex (coatomer).
What is a γ-tubulin ring complex (γ-TuRC)?
A complex of 7 γ-TuSC subunits in an overlapping spiral structure that acts as a template for microtubule nucleation.
What is SCAP (SREBP cleavage activation protein)?
A protein normally associated with SREBP in the ER membrane: at high [cholesterol], it has cholesterol bound, but at low [cholesterol], its lack of cholesterol triggers a conformational change that causes SCAP and SREBP to be transported to the Golgi, where SREBP is processed so that it can act as a transcription regulator for cholesterol synthesis.
Where does a nuclear pore complex get the energy to drive active transport of proteins into the nucleus?
Ran (Ras-related nuclear GTPase).
What fluorescent dye is used to stain DNA for visualization by fluorescence microscopy?
DAPI.
What is a t-SNARE?
A protein complex of 2 or 3 proteins found on a vesicle or target membrane that binds to a specific v-SNARE on a complementary target membrane or vesicle to form a trans-SNARE complex.
Why can the same cyclin-Cdk complex induce different effects at different times in the cell cycle?
The accessibility of some Cdk substrates changes during the cell cycle, so some targets are only available for phosphorylation at certain times.
What happens in M phase of the cell cycle?
Mitosis and cytokinesis.
In a nuclear pore complex, how do the fibrils attached to channel nucleoporins help with transport?
- Form a tangled mesh that makes it difficult for unwanted molecules to diffuse through
- Contain FG repeats as binding sites for nuclear receptors, which let them “hop” through the channel
What is monitored at the first cell-cycle checkpoint (start transition)?
Whether the environment is favourable for division.
What is a V-type proton pump?
An ATP pump activated by a single phosphorylation event to drive active transport of protons; used to regulate pH.
How does acetylcholine interact with GPCRs to reduce the rate and strength of heart muscle cell contraction?
- Acetylcholine binds GPCR
- GPCR activates Gi protein
- Gi-α inhibits adenylyl cyclase, reducing [cAMP]
- Gi-βγ opens K+ channels, making it more difficult for the cell to depolarize
When in the cell cycle is G1/S-cyclin active?
G1: the cyclin is made just after the start point and degraded at the beginning of S-phase.
What is a major difference between myosins (actin-associated motor proteins) and kinesins (microtubule-associated motor proteins)?
- Myosins:
- No ATP: tightly bound to filament
- ATP: released from filament
- Kinesins:
- No ATP: released from filament
- ATP: tightly bound to filament
Which two major classes of binding proteins regulate actin filament behaviours?
- Proteins that bind along the side of a filament
- Proteins that bind to the ends of a filament
How does M-Cdk participate in a positive feedback loop to promote activation of more M-Cdk?
M-Cdk activates CAK (Cdk-activating kinase) and inhibits Wee1 (a Cdk inhibitor).
What is DNA catenation?
The interlocking of DNA strands between sister chromatids to make them inseparable until cleaved and re-sealed by topoisomerase II.
True or false: Transporters can be involved in either active or passive transport.
True.
What happens to Rab-GTP once it has bound to its corresponding Rab effector on its vesicle’s target membrane?
It is hydrolyzed to Rab-GDP and becomes soluble in the cytosol.
What is the difference between homotypic and heterotypic fusion?
- Homotypic: membranes are from the same compartment, and SNARE interaction is symmetrical (v-SNAREs and t-SNAREs contributed by both membranes)
- Heterotypic: membranes are from different compartments, and SNARE interaction is asymmetrical (v-SNAREs on 1 membrane, t-SNAREs on the other)
How long does the S phase take in human cells?
About 12 hours (to fully replicate the human genome).
What happens to PI(4,5)P2 when it is cleaved by PLCβ?
It splits into diacylglycerol and inositol 1,4,5-trisphosphate (IP3); diacylglycerol stays bound to the plasma membrane, and IP3 becomes soluble in the cytosol, and both recruit further signalling molecules.
What is the key difference between FITC (fluorescent dye) and GFP/CFP/RFP/YFP (fluorescent proteins)?
FITC is added to a sample to stain all proteins non-selectively; fluorescent proteins must be added transgenically to proteins and can be used to tag specific proteins of interest for visualization.
What is clathrin?
A vesicular coat protein found on vesicles budding toward the cytosol from the plasma membrane or from the trans-Golgi membrane.
What is DAPI?
A fluorescent dye used to stain DNA for visualization by fluorescence microscopy.
What is a Na+-driven glucose symport?
A transporter that couples the active transport of glucose with the passive transport of Na+ into the cell
What is the role of PIP phosphatases in vesicle formation?
They remove a phosphate group from the PIP to which the coat assembly adaptor protein is bound, causing the adaptor protein and therefore the coat protein to dissociate from the completed vesicle.
What is the role of kinetochore microtubules in the mitotic spindle?
They attach to the chromatids at 1 end (plus) and to the centrosomes at the other (minus), and they pull the chromatids apart to either end of the cell during anaphase.
Which two transport proteins create and maintain the membrane potential in animal cells?
- Na+-K+ pump (P-type pump/transporter)
- K+ leak channel (ion channel)
What is p16?
A Cdk inhibitory protein (CKI) in mammalian cells that suppresses G1-Cdk activity in G1 (frequently inactivated in cancer).
Which actin homologue in bacterial cells helps during plasmid segregation?
ParM.
Which forms of Ran are found inside and outside of the nucleus?
- In the nucleus: Ran-GTP
- In the cytosol: Ran-GDP
What is Rab?
A monomeric GTPase that labels vesicles according to their membrane domain or organelle of origin and that helps vesicles tether and dock to their target membrane.
What is the simplest mechanism for intercellular communication in multicellular organisms?
Gap junctions.
What are Sec23/24 and Sec13/31?
2 sets of proteins that function as GAPs for Sar1-GTP in COPII-coated vesicles, promoting hydrolysis of Sar1-GTP to Sar1-GDP to trigger shedding of the protein coat once the vesicle is complete.
What is the role of interpolar microtubules in the mitotic spindle?
The minus ends are attached to the centrosomes at either end of the cell, and the plus ends of opposite MTs interact via kinesins to push and pull on the centrosomes until the entire apparatus is positioned correctly for anaphase.
What happens to transport vesicles once they have budded off from the ER?
They shed their COPII coats and fuse with one another to form vesicular tubular clusters, which then bud off transport vesicles of their own that have COPI coats.
Is [K+] higher inside or outside a cell? How is this maintained?
Inside (as maintained by Na+-K+ pumps and K+ leak channels).
What are the 3 main types of ATP pumps?
- P-type pumps
- ABC transporters
- V-type proton pumps
What triggers the opening of a mechanically gated ion channel?
High osmotic pressure causes the surrounding membrane to stretch and pull the channel pore open.
How does activation of two different G proteins by GPCRs result in the localized activation of Rac and Rho pathways at the front and back, respectively, of a moving lamellipodium?
- Rac: G protein activates PI 3-kinase to create membrane-bound PI(3,4,5)P3, which activates the Rac pathway at the front of the cell
- Rho: G protein dissociates into Gα & Gβγ, which can diffuse further down the cell to activate the Rho pathway at the back
(Rac inhibits Rho, so any Rho components activated at the front are immediately inactivated by Rac activity.)
What is a glycolipid?
A membrane lipid based on sphingosine, containing a single fatty acid tail (in addition to the fatty chain of sphingosine) and one or more sugars (where sphingolipids would have a phosphate group attached instead).
How does the use of COPII in ER vesicles and COPI in vesicular tubular cluster vesicles reduce the number of ER resident proteins accidentally leaving the ER?
ER resident proteins have a higher affinity for COPI than for COPII, so they are less likely to end up in vesicle budding from the ER and more likely to end up in retrograde transport vesicles from the clusters.
How do mechanically gated ion channels protect cells from rupturing due to osmotic pressure?
Osmotic pressure increases when cells take up water in response to increased ion concentration, so the opening of mechanically gated channels to move ions out of the cell results in water also leaving the cell to maintain the concentration gradient.
Which protein complexes are responsible for inserting proteins into the inner membrane of the mitochondria?
TIM22, TIM23, and OXA.
What fluorescent dye is used to stain proteins (non-selectively) for visualization by fluorescence microscopy?
FITC.
What does cohesin interact with in order to form a ring around sister chromatids to keep them together until anaphase?
2 Smc molecules.
What is a phosphoglyceride?
A type of membrane phospholipid based on a glycerol backbone, with 2 fatty acid tails, a phosphate group, and one of several possible head groups.
What is an inositol phospholipid?
A minor class of phospholipids that are phosphorylated to form phosphoinositides during cell signalling events.
What does TIC stand for?
Translocator of the inner chloroplast membrane.
What is separase?
An enzyme that cleaves cohesin so that sister chromatids can be separated into daughter cells during cell division.
What is an SRP receptor?
A receptor bound to the cytosolic face of the ER membrane that binds incoming SRPs, allowing protein translocators to bind to the SRP-bound protein’s signal sequence, which causes the associated ribosome to resume translation and the SRP to be released back into the cytosol.
What is a BAR domain protein?
A positively charged, crescent-shaped protein that helps to deform the plasma membrane via electrostatic interactions during clathrin coat assembly in vesicle formation.
Which vesicles are assembled with COPI or COPII protein coats?
Vesicles transported between the Golgi and the ER.
Where are centrosomes (MTOCs) usually located in the cell?
At the centre, with microtubules radiating outward (astral configuration).
How is cholesterol transported to cells?
Cholesterol aggregates within LDLs, which travel through the bloodstream and are bound by LDL receptors on the outer surface of cell membranes, which then trigger the assembly of clathrin-coated vesicles for import from the plasma membrane.
What are glutaraldehyde and osmium tetroxide used for in cell work?
To fix samples for electron microscopy.
What is the basic structure of a voltage-gated Na+ channel?
- 1 polypeptide made up of 4 similar domains, meeting in a ring to form a central channel
- Selectivity filter between α-helices
- Inactivation gate between domains 3 and 4
- Lateral portals connecting the central cavity to the hydrophobic portion of the membrane
Which proteins act as chaperones for precursor proteins destined for import into the mitochondria?
Cytosolic hsp70.
What sugar groups are found in the core region of the precursor oligosaccharide added to glycosylated proteins in the ER?
2 GlcNAc and 3 mannose.
What are the different responses to acetylcholine in skeletal muscle cells, heart pacemaker cells, and salivary gland cells?
- Skeletal muscle: muscle contraction
- Heart pacemaker: decreased firing rate
- Salivary gland: secretion
What is a K+ leak channel?
A channel protein that allows the diffusion of K+ across the plasma membrane to maintain the membrane potential of the cell by regulating cytosolic [K+].
How does the hsp70 chaperone protein interact with clathrin-coated vesicles?
It functions as an uncoating ATPase to help strip off the clathrin coat once the vesicle has detached from its donor membrane.
What happens to a nuclear import receptor once it has entered the nucleus through a nuclear pore complex?
Ran-GTP in the nucleus binds to the receptor, causing the receptor to dissociate from its cargo protein.
What is a sphingolipid?
A membrane lipid based on sphingosine, which incorporates a fatty chain tail capped by a hydroxyl group, with one additional fatty acid tail, a phosphate group, and one of several possible head groups.
Which 3 kinesins interact with microtubules to position the mitotic spindle?
- Kinesin-5 (where antiparallel microtubules meet; pushes poles outward)
- Kinesin-14 (pulls poles inward)
- Kinesin-4,10 (pulls DNA toward the centre of the spindle)
In the context of voltage-gated Na+ channels, what is the refractory period?
The time necessary for enough Na+ channels to recover from inactivation to support a new action potential.
What is galactocerebroside?
A glycolipid with a galactose residue at its head, found most commonly in myelin sheaths.
Which type of filament’s rapid assembly and disassembly drives the rapid movement of neutrophils?
Actin filaments.
What is FITC?
A fluorescent dye used to stain proteins non-selectively for visualization by fluorescence microscopy.
What are the 2 main signalling pathways that can follow from phosphorylation of PI(4,5)P2 by PI 3-kinase to PI(3,4,5)P3?
- PI(3,4,5)P3 can act as a docking site for intracellular signalling proteins
- PI(3,4,5)P3 can be cleaved by phospholipase C, with diacylglycerol remaining in the membrane and the rest of the molecule (IP3) diffusing into the cytosol, so each portion participates in different signalling activities
Which protein coat is found on vesicles transported from the ER to the Golgi?
COPII.
What two proteins form a complex to nucleate actin filaments in branching actin filament assemblies?
Arp2 and Arp3.
What is an ion channel?
A channel protein used to passively transport ions across a membrane.
What molecule activates condensin to trigger chromosome condensation during mitosis?
M-Cdk (by phosphorylation).
What does TOC stand for?
Translocator of the outer chloroplast membrane.
What is rhodamine B?
A fluorescently labelled antibody used to visualize specific proteins by fluorescence microscopy (emits red light).
What molecules are exchanged by the Ca2+ pumps found in the sarcoplasmic reticulum?
2H+ for 2Ca2+.
What is synaptotagmin?
A protein jutting out of a primed synaptic t-SNARE/v-SNARE-complexin complex that contains Ca2+ binding sites, responsible for activating the complex to allow release of neurotransmitters when a signal is received.
Which motor proteins are associated with actin filaments?
Myosins.
What is a kinetochore?
A protein that creates a binding area at the centromere of each chromatid for attachment of kinetochore microtubules during anaphase.
What is a polyprotein?
A single large protein that yields many copies of the same (smaller) protein when cleaved into fragments.
When does K+ stop diffusing out of a cell through a K+ leak channel?
When the electrical gradient opposes further K+ diffusion (even when the chemical gradient favours it).
How do CKIs inactivate cyclin-Cdk complexes?
By stimulating a large rearrangement in the structure of the Cdk active site, making it completely unable to bind its protein substrates.
Why is it important during mitosis that kinesin-14 walks in the opposite direction to all the other kinesins?
Kinesin-14 and kinesin-5 both interact with microtubules to position the mitotic spindle, and coordinated pushing and pulling allows more precise positioning of the centrosomes and DNA.
What is a protein signal sequence?
A sequence of amino acids in a protein that is recognized by sorting receptors and determines where in the cell the protein will be localized.
What is Rac-GTPase?
A monomeric GTPase that activates PAK and WASp family proteins, resulting in increased formation of branched actin webs and decreased formation of stress fibres in lamellipodia.
What is Smc?
A protein that binds in pairs via cohesin to form rings around sister chromatids, keeping them together until anaphase.
What is the basic structure of an individual clathrin protein?
A triskelion, with 3 heavy chains and 3 light chains.
What are axonemal (ciliary) dyneins?
Dyneins involved in beating of cilia and flagella.
What is Wee1 kinase?
A kinase that adds an inhibitory phosphate group to inactivate Cdk-cyclin complexes.
What does TIM stand for?
Translocator of the inner mitochondrial membrane.
How do morphogens determine the differentiation pathway of undifferentiated cells?
A morphogen concentration gradient: cells closer to the source have more receptors activated than cells further away, so cells at different positions along the gradient will react differently depending on how many receptors are activated in each.
What two layers of specificity are conferred by glyosyl transferases in the synthesis of glycosylated proteins?
- Different sugars require different transferases (each enzyme is specific to one sugar)
- Different cell types contain different transferases
True or false: Once a cell has reached its start point (restriction point) in the cell cycle, it is irreversibly committed to repeating the entire cycle (from G1 to S).
True (even if conditions become unfavourable for division).
What 3 domains are characteristic of a nuclear receptor protein?
- Ligand-binding domain
- DNA-binding domain
- Transcription-activating domain
What are WASp family proteins?
Proteins activated by Rac-GTP that activate Arp2 & Arp3 to stimulate nucleation of branched actin filaments, resulting in the formation of a branched actin web in lamellipodia.
What is a filapodium?
A 1-dimensional cell protrusion, created by a single actin filament extending from the cell.
How do motor proteins generate the energy needed to move along cytoskeletal filaments?
From repeated cycles of ATP hydrolysis.
What enhances the hydrolysis of GTP to GDP to inactivate G proteins?
RGS (regulator of G-protein signalling).
What is geminin?
A protein that inhibits Cdt1 in late G1 to prevent the formation of new preRCs.
What is PAK?
A kinase activated by Rac-GTP that inhibits MHC & MLCK, resulting in decreased myosin activity (and thus decreased stress fibre formation), and activates filamin, resulting in increased formation of branched actin webs in lamellipodia.
What is RFP (red fluorescent protein)?
A fluorescent protein used to transgenically label specific proteins for visualization by fluorescence microscopy (emits red light).
What is the difference between a voltage-gated Na+ channel in its closed state and in its inactive state?
An inactive channel is unable to reopen until after the membrane potential has returned to its initial negative value, while a closed channel is free to be reopened as soon as another action potential is initiated.
What does TOM stand for?
Translocator of the outer mitochondrial membrane.
Where are all transmembrane proteins made?
In the ER (except for transmembrane mitochondrial/chloroplast proteins).
Why do glycerols and cholesterol esters arrange themselves in monolayers rather than bilayers?
They have no hydrophilic portion.
What is a signal recognition particle (SRP)?
A protein-RNA complex that recognizes and binds emerging ER-import signal sequences of proteins being synthesized, pauses translation, and delivers the ribosome and protein to the ER membrane.
What is a transmitter-gated ion channel?
A type of ion channel found in cells on either side of a chemical synapse for highly selective transport of specific neurotransmitters.
What molecule is required for a nuclear import receptor to successfully deliver its protein cargo to the nucleus?
Ran-GTP.
What is the main functional difference between myosin II and myosin V?
Myosin V has a longer lever arm domain, so it can swing further with each “step”.
What is an inhibitory neurotransmitter?
A neurotransmitter that prevents membrane depolarization by causing Cl- or K+ channels to open, which makes it harder for Na+ channels to create an overall positive charge inside the cell.