Abstract
KRas4b is a small G-protein whose constitutively active oncogenic mutants are present in 90% of pancreatic cancers. Using fully post-translationally modified KRAS4b, we investigated the role of lipid identity in the recruitment of KRas4b to a membrane surface of defined composition. Application of a newly developed single frequency fluorescence anisotropy decay experiment to this system revealed that KRas4b has a significant binding preference for Nanodisc bilayers containing PIP2. We conducted molecular dynamics simulations to look for an origin of this specificity. In the case of membranes containing PIP2 the protein formed long-lived salt bridges with PIP2 head groups but not the monovalent DMPS, explaining the experimentally observed lipid specificity. Additionally, we report that PIP2 forms key contacts with Helix-4 on the catalytic domain of KRas4b that orient the protein in a manner expected to facilitate association with upstream and downstream signaling partners.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 351-355 |
| Number of pages | 5 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 487 |
| Issue number | 2 |
| DOIs | |
| State | Published - May 27 2017 |
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Keywords
- Cancer signaling
- KRas4b
- Lipid specificity
- Nanodisc
- PIP
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Interaction of KRas4b with anionic membranes : A special role for PIP2. / Gregory, Michael C.; McLean, Mark A.; Sligar, Stephen G.
In: Biochemical and Biophysical Research Communications, Vol. 487, No. 2, 27.05.2017, p. 351-355.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interaction of KRas4b with anionic membranes
T2 - A special role for PIP2
AU - Gregory, Michael C.
AU - McLean, Mark A.
AU - Sligar, Stephen G.
PY - 2017/5/27
Y1 - 2017/5/27
N2 - KRas4b is a small G-protein whose constitutively active oncogenic mutants are present in 90% of pancreatic cancers. Using fully post-translationally modified KRAS4b, we investigated the role of lipid identity in the recruitment of KRas4b to a membrane surface of defined composition. Application of a newly developed single frequency fluorescence anisotropy decay experiment to this system revealed that KRas4b has a significant binding preference for Nanodisc bilayers containing PIP2. We conducted molecular dynamics simulations to look for an origin of this specificity. In the case of membranes containing PIP2 the protein formed long-lived salt bridges with PIP2 head groups but not the monovalent DMPS, explaining the experimentally observed lipid specificity. Additionally, we report that PIP2 forms key contacts with Helix-4 on the catalytic domain of KRas4b that orient the protein in a manner expected to facilitate association with upstream and downstream signaling partners.
AB - KRas4b is a small G-protein whose constitutively active oncogenic mutants are present in 90% of pancreatic cancers. Using fully post-translationally modified KRAS4b, we investigated the role of lipid identity in the recruitment of KRas4b to a membrane surface of defined composition. Application of a newly developed single frequency fluorescence anisotropy decay experiment to this system revealed that KRas4b has a significant binding preference for Nanodisc bilayers containing PIP2. We conducted molecular dynamics simulations to look for an origin of this specificity. In the case of membranes containing PIP2 the protein formed long-lived salt bridges with PIP2 head groups but not the monovalent DMPS, explaining the experimentally observed lipid specificity. Additionally, we report that PIP2 forms key contacts with Helix-4 on the catalytic domain of KRas4b that orient the protein in a manner expected to facilitate association with upstream and downstream signaling partners.
KW - Cancer signaling
KW - KRas4b
KW - Lipid specificity
KW - Nanodisc
KW - PIP
UR - http://www.scopus.com/inward/record.url?scp=85017450313&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85017450313&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2017.04.063
DO - 10.1016/j.bbrc.2017.04.063
M3 - Article
C2 - 28412347
AN - SCOPUS:85017450313
VL - 487
SP - 351
EP - 355
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -