TY - JOUR
T1 - Use of a two-hybrid system to investigate molecular interactions of GAP-43
AU - Chao, Steven
AU - Benowitz, Larry I.
AU - Krainc, Dimitri
AU - Irwin, Nina
N1 - Funding Information:
This work was supported by the National Eye Institute (NIH EY 05690 to L.B.) and the Boston Neurosurgical Foundation. We are grateful to Dr. Roger Brent (Massachusetts General Hospital) for advice regarding the interaction trap and for yeast strains and plasmids.
PY - 1996/9/1
Y1 - 1996/9/1
N2 - We used the 'interaction trap' (two-hybrid system) to identify polypeptides that interact with the neuronal phosphoprotein, GAP-43, in an intracellular environment. GAP-43 (neuromodulin, B-50, F1), a protein kinase C (PKC) substrate important for the growth and plasticity of neuronal connections, has been implicated in vitro in several signal transduction pathways. In the yeast-based cloning system, the only strong interaction that was detected was between GAP-43 and the calcium effector protein, calmodulin (CaM). PKC phosphorylates GAP-43 on serine 41. When we changed this serine to an aspartate residue to mimic constitutive phosphorylation, the interaction with CaM was blocked. Surprisingly, the N-terminal third of GAP-43 alone bound CaM more strongly than did intact GAP-43, suggesting that the protein's C-terminus may play a role in modulating the interaction with CaM. These results, along with other recent findings, suggest a novel role for the interaction between GAP-43 and CaM.
AB - We used the 'interaction trap' (two-hybrid system) to identify polypeptides that interact with the neuronal phosphoprotein, GAP-43, in an intracellular environment. GAP-43 (neuromodulin, B-50, F1), a protein kinase C (PKC) substrate important for the growth and plasticity of neuronal connections, has been implicated in vitro in several signal transduction pathways. In the yeast-based cloning system, the only strong interaction that was detected was between GAP-43 and the calcium effector protein, calmodulin (CaM). PKC phosphorylates GAP-43 on serine 41. When we changed this serine to an aspartate residue to mimic constitutive phosphorylation, the interaction with CaM was blocked. Surprisingly, the N-terminal third of GAP-43 alone bound CaM more strongly than did intact GAP-43, suggesting that the protein's C-terminus may play a role in modulating the interaction with CaM. These results, along with other recent findings, suggest a novel role for the interaction between GAP-43 and CaM.
KW - Calmodulin
KW - Development
KW - GAP-43
KW - Growth cone
KW - Phosphoprotein
KW - Protein kinase C
KW - Two-hybrid system
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U2 - 10.1016/0169-328X(96)00049-6
DO - 10.1016/0169-328X(96)00049-6
M3 - Article
C2 - 8872303
AN - SCOPUS:0007465152
SN - 0169-328X
VL - 40
SP - 195
EP - 202
JO - Molecular Brain Research
JF - Molecular Brain Research
IS - 2
ER -