Abstract
CRM1 (Exportin1/XPO1) exports hundreds of broadly functioning protein cargoes out of the cell nucleus by binding to their classical nuclear export signals (NESs). The 8- to 15-amino-acid-long NESs contain four to five hydrophobic residues and are highly diverse in both sequence and CRM1-bound structure. Here we examine the relationship between nuclear export activities of 24 different NES peptides in cells and their CRM1-NES affinities. We found that binding affinity and nuclear export activity are linearly correlated for NESs with dissociation constants (Kds) between tens of nanomolar to tens of micromolar. NESs with Kds outside this range have significantly reduced nuclear export activities. These include two unusually tight-binding peptides, one from the nonstructural protein 2 of murine minute virus (MVM NS2) and the other a mutant of the protein kinase A inhibitor (PKI) NES. The crystal structure of CRM1-bound MVM NS2NES suggests that extraordinarily tight CRM1 binding arises from intramolecular contacts within the NES that likely stabilizes the CRM1-bound conformation in free peptides. This mechanistic understanding led to the design of two novel peptide inhibitors that bind CRM1 with picomolar affinity.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2037-2044 |
| Number of pages | 8 |
| Journal | Molecular Biology of the Cell |
| Volume | 29 |
| Issue number | 17 |
| DOIs | |
| State | Published - Aug 15 2018 |
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ASJC Scopus subject areas
- Molecular Biology
- Cell Biology
Cite this
Correlation of CRM1-NES affinity with nuclear export activity. / Fu, Szu Chin; Fung, Ho Yee Joyce; Caǧatay, Tolga; Baumhardt, Jordan; Chook, Yuh Min.
In: Molecular Biology of the Cell, Vol. 29, No. 17, 15.08.2018, p. 2037-2044.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Correlation of CRM1-NES affinity with nuclear export activity
AU - Fu, Szu Chin
AU - Fung, Ho Yee Joyce
AU - Caǧatay, Tolga
AU - Baumhardt, Jordan
AU - Chook, Yuh Min
PY - 2018/8/15
Y1 - 2018/8/15
N2 - CRM1 (Exportin1/XPO1) exports hundreds of broadly functioning protein cargoes out of the cell nucleus by binding to their classical nuclear export signals (NESs). The 8- to 15-amino-acid-long NESs contain four to five hydrophobic residues and are highly diverse in both sequence and CRM1-bound structure. Here we examine the relationship between nuclear export activities of 24 different NES peptides in cells and their CRM1-NES affinities. We found that binding affinity and nuclear export activity are linearly correlated for NESs with dissociation constants (Kds) between tens of nanomolar to tens of micromolar. NESs with Kds outside this range have significantly reduced nuclear export activities. These include two unusually tight-binding peptides, one from the nonstructural protein 2 of murine minute virus (MVM NS2) and the other a mutant of the protein kinase A inhibitor (PKI) NES. The crystal structure of CRM1-bound MVM NS2NES suggests that extraordinarily tight CRM1 binding arises from intramolecular contacts within the NES that likely stabilizes the CRM1-bound conformation in free peptides. This mechanistic understanding led to the design of two novel peptide inhibitors that bind CRM1 with picomolar affinity.
AB - CRM1 (Exportin1/XPO1) exports hundreds of broadly functioning protein cargoes out of the cell nucleus by binding to their classical nuclear export signals (NESs). The 8- to 15-amino-acid-long NESs contain four to five hydrophobic residues and are highly diverse in both sequence and CRM1-bound structure. Here we examine the relationship between nuclear export activities of 24 different NES peptides in cells and their CRM1-NES affinities. We found that binding affinity and nuclear export activity are linearly correlated for NESs with dissociation constants (Kds) between tens of nanomolar to tens of micromolar. NESs with Kds outside this range have significantly reduced nuclear export activities. These include two unusually tight-binding peptides, one from the nonstructural protein 2 of murine minute virus (MVM NS2) and the other a mutant of the protein kinase A inhibitor (PKI) NES. The crystal structure of CRM1-bound MVM NS2NES suggests that extraordinarily tight CRM1 binding arises from intramolecular contacts within the NES that likely stabilizes the CRM1-bound conformation in free peptides. This mechanistic understanding led to the design of two novel peptide inhibitors that bind CRM1 with picomolar affinity.
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UR - http://www.scopus.com/inward/citedby.url?scp=85051556232&partnerID=8YFLogxK
U2 - 10.1091/mbc.E18-02-0096
DO - 10.1091/mbc.E18-02-0096
M3 - Article
C2 - 29927350
AN - SCOPUS:85051556232
VL - 29
SP - 2037
EP - 2044
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 17
ER -