|
PKC Constructs |
PKC-HA |
|
Vectors |
pHACE or pHACB |
|
PKC Mutants |
WT |
DN |
ΔPS |
ΔNPS |
CAT |
CAT-KR |
|
Mutation (Diagram) |
wild type |
dominant negative (kinase inactive) |
pseudosubstrate sequence deletion |
N-terminal pseudosubstrate domain deletion |
catalytic domain |
catalytic domain mutant (kinase inactive) |
|
PKCα |
2-672 |
2-672 (K368R) |
2-672 |
30-672 |
326-672 |
326-672 (K368R) |
|
(Δ21-29) |
|
PKCß1 |
2-671 |
2-671 (K371R) |
N/A |
30-671 |
329-671 |
N/A |
|
PKCß2 |
2-673 |
2-673 (K371R) |
N/A |
30-673 |
329-673 |
N/A |
|
PKCγ |
2-697 |
2-697 (K380R) |
N/A |
29-697 |
338-697 |
N/A |
|
PKCδ |
2-674 |
2-674 (K376R) |
2-674 |
152-674 |
334-674 |
334-674 (K376R) |
|
(Δ143-151) |
|
PKCε |
2-732 |
2-732 (K437R) |
2-732 |
164-732 |
395-737 |
395-737 (K437R) |
|
(Δ155-163) |
|
PKCη |
2-683 |
2-683 (K384R) |
N/A |
166-683 |
342-683 |
N/A |
|
PKCζ |
2-592 |
2-592 (K281M) |
2-592 |
204-592 |
239-592 |
239-592 (K281R) |
|
(Δ195-203) |
|
PKCι |
2-587 |
2-587 (K273R) |
N/A |
125-587 |
232-587 |
N/A |
|
Data |
Structures of the PKC mutants |
Structure of pHACE-PKC constructs |
|
Expression level in COS-7 cells |
Coding
sequences of the PKC mutants |
|
Western
blot data |
Kinase assay data |
|
Immunofluorescence microscopy |
Test
digestion protocol |
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WT: wild type |
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DN(KR): dominant negative (kinase inactive) mutant |
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ΔPS: pseudosubstrate sequence deletion mutant (constitutively active) |
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ΔNPS: N-terminal pseudosubstrate domain deletion mutant (constitutively
active) |
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CAT: catalytic domain mutant (constitutively active) |
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CAT-KR: catalytic domain mutant (kinase inactive) |
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