f*****Y 发帖数: 20 | 1 请问各位达人:
需要做site directed mutagenesis to mimic tyrosine phosphorylation,
应该把tyrosine变成什么residue?
谢谢! | b******y 发帖数: 627 | 2 Glutamate.
But keep in mind: sometimes it does mimic functional and in equal number of
cases, it does not. | l****e 发帖数: 157 | 3 for in vivo or in vitro studies?
【在 f*****Y 的大作中提到】 : 请问各位达人: : 需要做site directed mutagenesis to mimic tyrosine phosphorylation, : 应该把tyrosine变成什么residue? : 谢谢!
| f*****Y 发帖数: 20 | 4 What is the major difference of using such a mutant in in vivo or in vitro
studies?
I hope it can be used for both.
Thank you both for the reply. | b******y 发帖数: 627 | 5 If the phosphorylation event is for disruption of an otherwise interactions,
YtoE works. However, if the phosphorylation event is to recruit a SH2 or
PTB containing binding partner, most likely this substitution will not work.
Of course, there are other scenarios this should or should not work.
good luck. | l**********1 发帖数: 5204 | 6 Here you go
Deramaudt TB et al. (2011)
FAK phosphorylation at Tyr-925 regulates cross-talk between focal adhesion
turnover and cell protrusion.
Mol Biol Cell. 22: 964-75.
Abstract
Cell migration is a highly complex process that requires the coordinated
formation of membrane protrusion and focal adhesions (FAs). Focal adhesion
kinase (FAK), a major signaling component of FAs, is involved in the
disassembly process of FAs through phosphorylation and dephosphorylation of
its tyrosine residues, but the role of such phosphorylations in nascent FA
formation and turnover near the cell front and in cell protrusion is less
well understood. In the present study, we demonstrate that, depending on the
phosphorylation status of Tyr-925 residue, FAK modulates cell migration via
two specific mechanisms. FAK⁻/⁻ mouse embryonic fibroblasts (
MEFs) expressing nonphosphorylatable Y925F-FAK show increased interactions
between FAK and unphosphorylated paxillin, which lead to FA stabilization
and thus decreased FA turnover and reduced cell migration. Conversely, MEFs
expressing phosphomimetic Y925E-FAK display unchanged FA disassembly rates,
show increase in phosphorylated paxillin in FAs, and exhibit increased
formation of nascent FAs at the cell leading edges. Moreover, Y925E-FAK
cells present enhanced cell protrusion together with activation of the p130(
CAS)/Dock180/Rac1 signaling pathway. Together, our results demonstrate that
phosphorylation of FAK at Tyr-925 is required for FAK-mediated cell
migration and cell protrusion.
link:
//www.ncbi.nlm.nih.gov/pubmed/21289086
from
or JBC 2004 one article:
//www.ncbi.nlm.nih.gov/pubmed/15117950
and cited this paper all PubMed records:
//www.ncbi.nlm.nih.gov/pubmed?linkname=pubmed_pubmed_citedin&from_uid=
15117950
【在 f*****Y 的大作中提到】 : What is the major difference of using such a mutant in in vivo or in vitro : studies? : I hope it can be used for both. : Thank you both for the reply.
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