In the last several classes, we've focused on receptors and signaling pathways. Today we look
at the last link in that chain, the transcription factor phosphorylation that leads to actual changes
in gene expression and cell behavior
Cue > Signals > Responses
We talked about signals the last 2 classes. We'll talk about cues for the following two, and
about responses (transcriptional, mostly) during the last 2 classes in this module.
Lecture 3 Notes:
Co Immunoprecipitation (CoIP) and Mass Spectrometry (MS)
CoIP, as the name implies, is about precipitating a particular protein by attaching it to
antibodies, and then looking to see what else gets dragged down with it.
Lecture 1 Notes:
The MAPK Case
In vivo, things are a tad more complicated. There's a scaffold protein in yeast called Sterile 5,
which holds together all the elements of the cascade in the right orientation and proximity. This
increases the overall rate o
Lecture 2 Notes: Titration Analysis
Today we'll talk about some experimental techniques that can give us the useful
parameters for our models.
1. Attach protein to plate
2. Add fluorescently labeled
ligand 3. Rinse off
Lecture 6 Notes: Constitutive Kinases
We were talking about kinases and their activation loops.
Let's now go into greater detail of how kinases work and how they fit into signaling cascades.
We'll also tie it to EGFR.
EGFR normally exists at abo
Lecture 8 Notes: FACS
But how do we measure the amount of surface receptors? We could do FACS with antiEGFR
antibodies tagged with fluorophores. This is great, and measures just the surface receptors.
Most people for a long time just did Western blots, wh
Regulation 1. Simple
4. Local concentration of
signal 5. Different amounts
6. Different amounts/a
Lecture 5 Notes:
Last time we talked about EGFR and the importance of that signaling pathway. It leads to cell
proliferation, so all manner of pharmaceutical companies are trying to inhibit it to treat cancer.
We'll try to underst
Lecture 4 Notes:
We get Kd from multiple [L]o
We get koff from dissociation, which gives us kon because we know
Kd. We get Kon and koff from the association phase.
Limitations of SPR
Binding of protein/ligan