Cell Bio Final Flashcards
Gustavo Silva
Oxidative stress in cells and uses bakers yeast to study the mechanisms
Cellular oxidative stress is characterized by an imbalance between reactive oxygen species production and intracellular antioxidant defense, leading to potential damage
Three major types of receptors
GPCR (G-protein-coupled receptors
Enzyme-Coupled Receptors
Ion-Channel- Coupled Receptors
How is signaling managed?
Via protein phosphorylation/dephosphorylation (post translational modification) OR GTP nucleotide binding & hydrolysis
Dephosphorylation
phosphatase: removing phosphate→ inactivates
Phosphorylation
kinase: activates
GEF
GDP to GTP to turn on
GAP
removing phosphate to turn off
Signal by protien phosphorylation
ATP to ADP using protien kinase to add phosphate and turn on
then uses protien phosphatase to remove phosphate turning off signal
Signaling by GTP binding protien
GDP (off) to GTP (on) using GTP binding turning on signal
then using GTP hydrolysis removing the phosphate and turning off signal
Endocrine
Hormones → bloodstream →body.
Paracrine
signals →neighbors (cell A acts on cell B)
Autocrine
cell signals itself “auto”
Neuronal
electrical signals →nerve cell axon→nerve terminal→ neurotransmitter release→target cells
Contact-dependent
cells must physically interact to signal
Ligand/Receptor Interaction
Ligand (signal molecule)
- hormone/drug/ neurotransmitter
- Causes conformational change
Receptor: ligand binds
Activation of Heterotrimeric G-protein
- The alpha dissociates from the the Beta Gamma (Y)
- GTP binds to alpha (on signal)
Heterodimers can form due to sharing a common ligand
Signal Transduction
when a cell responds to the ligand-receptor binding
Signal Amplification
Amplifying signals using different enzymes
Also used second messengers, ex: calcium
RTK (receptor tyrosine kinase)
type of cell surface receptors
Dephosphorylation/phosphatase: removing phosphate→ inactivates
Phosphorylation/kinase: activates
GPCR (G-protein coupled receptor) and RTK receptors
Immediate Targets
Immediate: less than seconds to minutes
Fast
Signal binds to receptor→ intracellular IN CYTOPLASM altered protein function→altered cytoplasmic machinery→altered cell behavior
Peptide hormones (hydrophilic)
Longer Targets
Longer: min to hours
Slow
Signal binds to receptor→ IN NUCLEUS (DNA + RNA) → altered protein synthesis→altered cytoplasmic machinery→altered cell behavior→control gene expression
Steroid hormones (hydrophobic)
Steroid hormone – can be initially cytosolic or nuclear but bind ligand (ligand passes through cell membranes) and makes receptor active in controlling gene expression
GPCR – signaling through cAMP
cAMP is formed from ATP by adenylate cyclase
Inactivated by hydrolysis to AMP by PDE
GPCR – signaling through PLC (phospholipase C) → IP3 & DAG second messengers
DAG attaches to plasma membrane and recruits protein kinase C (direct effect)
IP3 diffuses to the ER and is bound the IP3 receptor
The IP3 receptor serves as a calcium channel and releases calcium from the ER
- Calcium binds to protein kinase C and others and activates it
IP3 indirectly acts on PKC via Ca+2 ions
RTK – signal to Ras and MAPK
Receptor tyrosine kinases respond to DIMERIC signals- these serve to dimerize & activate the receptor
RTK targets Ras protein & MAPK cascade
RTK type of cell surface receptors - dephosphorylation and phosphorylation - tyrosine residue
Ras
Ras proteins is a cellular oncogene
GTP/GDP binding
Changes confirmation due to one or the other nucleotide being bound cause changes in target proteins it binds or controls
RAS activates the MAPK which transmits signals downstream resulting in transcription
MAPK
MAPK: mitogen activated protein (MAP) kinase→ transcription
Kinase cascade moves this signal
Tor
Tor is a kinase: a major regulator of metabolism and growth in all eukaryotes
Downstream target of RTKs
Notch-Delta signaling
Notch processing via protease cleavage
Upregulation and downregulation
Differentiation in the cell process, can delay the cells
Delta is ligand (signal)
Notch is receptor
The delta signal protein will bind to the delta receptor notch
When they bind, a cleaved notch tail migrates to the nucleus→ transcription of notch-responsive genes
Calmodulin
Alterations in Ca+2 are often “read” by calmodulin or calmodulin kinase
Calmodulin is a calcium binding protein that mediates calcium regulation
Activated by intracellular calcium
Mutant Ras
Mutant Ras that is always “on” can be useful
“Dominant active” or “gain of function”
Does gene X or gene Y act upstream of Ras?
Dominant active rescues, so Ras should come after X, not before, otherwise loss of X block effect of dominant (always on) Ras
The opposite situation when the mutant gene Y functions downstream of Ras (you don’t know this before you do the experiment. What about the loss of Y?
Dominant active does not rescue loss of Ym so Ras should come before Y, not after, otherwise loss of Y would not block effect of dominant (alway on) Ras
Intracellular signals and second messengers
Levels of second messengers controlled during signaling enzymatic reactions or opening of ion channels to ensure that they are highly amplified
Second messengers are generated within the cells as a downstream step in signal transduction
Binding GTP to the G protein leads to
dissociation of the G-protein from the receptor
Activation of G protein
Steps A to D
A. Resting heterotrimer state of the G protein. GDP is bound to the alpha subunit in resting state and receptor is not associated with G protein.
B. Signal molecule (Ligand) binding to G protein receptor causes G protein to become physically associated with G protein receptor which then causes a conformational change that results in a reversal of the G proteins relative affinities for GDP/GTP.
C. GTP binding cause heterotrimer to disassociate into two activated subunits alpha and beta gamma.
D. beta gamma subunits opens the K+ (potassium) channel
Inactivation of G protein
Steps
Extracellular Space
Cytosol activated by beta gamma complex
Activated alpha subunit acts like GTPase - hydrolysis of GTP by the alpha inactivates and causes it to dissociate from the target protein
Inactive alpha reassembles with beta gamma complex to reform an inactive G protein
RTK can be down regulated by
RTK can be down regulated by protein tyrosine phosphatases dephosphorylates the RTKs or by endocytosis and lysosomal targeting of the receptor from the endosome the receptor many also recycle back to the plasma membrane
The axon of a neuron transmits an electrical signal from its cell body to its synapse where it releases a neurotransmitter agent onto its target cell. What of the following is used to describe the general process that changes the electrical signal to a chemical signal?
Transduction
Which of the following would explain the net inward movement of solute X into the cell?
Concentration of X higher outside than inside
An inward directed energy requiring a pump mechanism located in the plasmalemma
True or False: Membranes can be permeable to some small uncharged yet polar molecules?
True
For most animal cells the distribution of Na+ ions across the plasma membrane is such that
Na+ is higher outside the cell
Na (sodium)
Na+ ions are maintained at a higher level outside than inside the cell largely by the activity of what?
Na+/K+ ATPase pump
True or False: Channel proteins can perform active transport
False