Abstract:
:To explore the structural basis required for the holoenzyme formation of cAMP-dependent protein kinase, we have prepared rabbit anti-peptide antibodies that can block the holoenzyme formation without affecting the catalytic activity of the enzyme. The antibodies were raised against a specific site in the catalytic (C)-subunit, termed IDA (Inter-DFG-APE) region, which lies between the kinase subdomains VII and VIII. Although the C-subunit immunoprecipitated with anti-IDA antibodies could not form a stable complex with regulatory (R)-subunit, it was still susceptible to inhibition by the R-subunit or by PKI, a specific inhibitor peptide containing a pseudosubstrate site. These results indicate that there exists an IDA region-mediated interaction between the R- and C-subunits, which is distinct from that mediated through the substrate site and substrate binding site. In accordance with this idea, association of synthetic IDA peptides with the R-subunit was directly demonstrated by resonance mirror analysis. The calculated association constants of IDA peptides were high enough to suggest a possible involvement of the IDA region in the initial step of holoenzyme formation.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Sahara S,Sato K,Kaise H,Mori K,Sato A,Aoto M,Tokmakov AA,Fukami Ydoi
10.1016/0014-5793(96)00302-xsubject
Has Abstractpub_date
1996-04-15 00:00:00pages
138-42issue
2eissn
0014-5793issn
1873-3468pii
001457939600302Xjournal_volume
384pub_type
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