L9 - Beta Subunits Flashcards
What are beta subunits
Small proteins that may be cytosolic or integral membrane proteins
They interact with ion channels
How do beta subunits interact with ion channels
Through impacting on their trafficking and open prob
What is the KCNQ1 channel regulated by
KCNE
KCNQ1 is the
Alpha subunit
Normal function of an alpha subunit is only seen when
The beta subunit is alo present
How many members of the KCNE family
KCNE1-5
How many amnio acids in the E family
103-177
How many TMDs does the E family have
1 transmembrane domain
Describe the expression of KNCNE1-5
Expressed widely in excitable cells - implicated in long QT syndrome
What do the properties of Q1 depend on
The beta subunits that are regulating it
To investigate the effects of E1 on Q1 what experiements were performed
Two electrode vltage clamp on xeonopus oocytes expressing cRNA encoding Q1 and Q1andE1
Describe the differences in current seen when Q1 was expressed alone to when Q1 was expressed with E1
E1 enhances function of Q1
See a shift in time dependence - currents reach a steady state earlier when E1 present
Where is KCNE1 expreesed
In the proximal tubule
Describe the localisation of KCNE1
How does this relate to KNCQ1 expression
In rings - apical membrane of proximal tubule cells of the mouse
Some overlap in E1/Q1 but also many areas that are E1+ and Q1-
What is the function of the proximal tubule
Early-mid important for the reabsorpition of glucose
Creates a driving force for Na uptake (also for glucose through the Na/glucose transporter
Describe the codnitoins for a clearance study
Anesthtised mouse on heated pad
Cannulate; carotid artery, jugular vein and bladder
Describe why the mouse is on a heated pad
To maintain a normal body temperature
Describe why the carotid pad needs to be cannulated
Ensure there is a viable BP
Can use this to determine the depth of anesthesia
Can take a blood sample at the end
Why is the jugular vein cannulated
To replace fluid
Why is the bladder cannulated
To measure volume per unit time - composition of the urine
Describe the plasma Na/Cl in the WT and E1 KO
Unchanged
Describe the plasma glucose levels in the WT vs E1 KO
Why might this not be accurate?
Plasma glucose is higher in the WT than E1 KO
Initially plasma glucose much higher than it should have been
What was seen RE GFR in the WT and E1 KO
GFR is unchanged in the wildtype and E1 KO