e*******c
发帖数: 1479 | 1 各位同仁，AKT308的磷酸化调控对于473的磷酸化有何作用？ PI3K通过PDK1调控着308
，然后如何作用于473的？auto磷酸化的作用？
我们发现一个非PI3K依赖性的473变化，该如何解释呢？谢谢 |
j********r
发帖数: 156 | 2 Generally, the kinases for T308 and S473 of Akt are PDK1 and mTOR complex 2
respectively. PI3K causes the production of PIP3 on the membrane, which then
binds to both Akt and PDK1 (which is assumed to be constitutively active).
So far there seems no direct evidence as to how mTORC2 is regulated. However
, inhibition of PI3K in most cases will lead to the suppression of
phosphorylation at S473.
Just curious, what is the evidence for PI3K-independent S473 phosphorylation
in your experiment? PI3K have different catalytic and regulatory subunits,
thus targeting PI3K genetically or via small molecules might not result in
loss of its kinase activity. |
w***a
发帖数: 4361 | 3 473是mTOR磷酸化的吧？本来跟PI3K关系就不直接啊
308
【在 e*******c 的大作中提到】
: 各位同仁，AKT308的磷酸化调控对于473的磷酸化有何作用？ PI3K通过PDK1调控着308
: ，然后如何作用于473的？auto磷酸化的作用？
: 我们发现一个非PI3K依赖性的473变化，该如何解释呢？谢谢
|
e*******c
发帖数: 1479 | 4 我同意上位的观点，就是有可能在PI3K缺陷细胞或小分子抑制剂的情况下并没有完全
block其活性，我用了一个试验是在PI3K的p85缺陷细胞中检测到仍然有AKT473的调节，
但是否一定说明我这个调节是PI3K-independent的还为时过早。
为伊消的人憔悴。 |
e*******c
发帖数: 1479 | 5 我同意上位的观点，就是有可能在PI3K缺陷细胞或小分子抑制剂的情况下并没有完全
block其活性，我用了一个试验是在PI3K的p85缺陷细胞中检测到仍然有AKT473的调节，
但是否一定说明我这个调节是PI3K-independent的还为时过早。
为伊消的人憔悴。 |
j********r
发帖数: 156 | 6 That seems more complex. The loss of PI3KR1 has been shown to cause
different or even opposite effects on downstream signaling. I would
recommend the use the p110 alpha or beta-null cells or specific inhibitor (
such as PIK75, TGX221) for your purpose. But it might not worth the effort
unless there is a potential to find out a new regulatory pathway or new
protein. |
e*******c
发帖数: 1479 | 7 in which kind of condition the lost of p85 has opposite effect? especially
for p-AKT? i thought about p110, and anyone know the K/O cells avaiable? |
s********n
发帖数: 248 | 8 你使用的那个p85 isoform缺陷啊？
【在 e*******c 的大作中提到】
: 我同意上位的观点，就是有可能在PI3K缺陷细胞或小分子抑制剂的情况下并没有完全
: block其活性，我用了一个试验是在PI3K的p85缺陷细胞中检测到仍然有AKT473的调节，
: 但是否一定说明我这个调节是PI3K-independent的还为时过早。
: 为伊消的人憔悴。
|
