d*x
发帖数: 51 | 1 I found my protein is phosphorylated and stabilized by phosphorylation,
which enhances its occupancy of target gene promoter, but decreases its
activity. Does anyone have any clue to explain this phenomenon?Thanks |
d*x
发帖数: 51 | 2
【在 d*x 的大作中提到】
: I found my protein is phosphorylated and stabilized by phosphorylation,
: which enhances its occupancy of target gene promoter, but decreases its
: activity. Does anyone have any clue to explain this phenomenon?Thanks
|
d*x
发帖数: 51 | 3 conformation cahnge? any ideas |
e****s
发帖数: 1125 | 4 Your information is too general. I believe that it is hard for anyone to
give any meaningful suggestions.
It is normal that phosphorylation changes protein's conformation.
I don't know much about protein-DNA interaction. But the enhancement of
protein's binding with target DNA sequence by phosphorylation should not be
a big surprise.
Regarding stability and activity of the protein, have no idea.
What made you think that the phosphorylation stabilizes the protein? |
d*x
发帖数: 51 | 5 Thanks, Emases,
I did the half life of the protein, the phosphoralated protein more stable. |
e****s
发帖数: 1125 | 6 Apparently, you have purified protein. Then why not trying to detect the
conformational change?
Limited proteolysis, fluorescence or other biophysical methods.
【在 d*x 的大作中提到】
: Thanks, Emases,
: I did the half life of the protein, the phosphoralated protein more stable.
|
v**********m
发帖数: 5516 | 7 activator conformation -----phosphorylated------> inhibitor conformation.
【在 d*x 的大作中提到】
: I found my protein is phosphorylated and stabilized by phosphorylation,
: which enhances its occupancy of target gene promoter, but decreases its
: activity. Does anyone have any clue to explain this phenomenon?Thanks
|
d*x
发帖数: 51 | 8 I found my protein is phosphorylated and stabilized by phosphorylation,
which enhances its occupancy of target gene promoter, but decreases its
activity. Does anyone have any clue to explain this phenomenon?Thanks |
d*x
发帖数: 51 | 9
【在 d*x 的大作中提到】
: I found my protein is phosphorylated and stabilized by phosphorylation,
: which enhances its occupancy of target gene promoter, but decreases its
: activity. Does anyone have any clue to explain this phenomenon?Thanks
|
d*x
发帖数: 51 | 10 conformation cahnge? any ideas |
|
|
e****s
发帖数: 1125 | 11 Your information is too general. I believe that it is hard for anyone to
give any meaningful suggestions.
It is normal that phosphorylation changes protein's conformation.
I don't know much about protein-DNA interaction. But the enhancement of
protein's binding with target DNA sequence by phosphorylation should not be
a big surprise.
Regarding stability and activity of the protein, have no idea.
What made you think that the phosphorylation stabilizes the protein? |
d*x
发帖数: 51 | 12 Thanks, Emases,
I did the half life of the protein, the phosphoralated protein more stable. |
e****s
发帖数: 1125 | 13 Apparently, you have purified protein. Then why not trying to detect the
conformational change?
Limited proteolysis, fluorescence or other biophysical methods.
【在 d*x 的大作中提到】
: Thanks, Emases,
: I did the half life of the protein, the phosphoralated protein more stable.
|
v**********m
发帖数: 5516 | 14 activator conformation -----phosphorylated------> inhibitor conformation.
【在 d*x 的大作中提到】
: I found my protein is phosphorylated and stabilized by phosphorylation,
: which enhances its occupancy of target gene promoter, but decreases its
: activity. Does anyone have any clue to explain this phenomenon?Thanks
|
l**********e
发帖数: 146 | 15 DNA binding增加了多少？活性减少了多少？
从你提供的信息看，这个磷酸化有两个作用：
（1）磷酸化->稳定蛋白—>DNA binding增加，那么DNA binding增加就是由于蛋白多了
导致的（相当于overexpression）
（2）DNA binding 之后，转录活性降低，说明磷酸化抑制了transactivation
(1)和（2）不矛盾，结论应该是磷酸化的overall effect还是抑制转录活性 |
