Allosteric activation of apicomplexan calcium-dependent protein kinases.

Abstract:

:Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7-kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes.

authors

Ingram JR,Knockenhauer KE,Markus BM,Mandelbaum J,Ramek A,Shan Y,Shaw DE,Schwartz TU,Ploegh HL,Lourido S

doi

10.1073/pnas.1505914112

subject

Has Abstract

pub_date

2015-09-08 00:00:00

pages

E4975-84

issue

36

eissn

0027-8424

issn

1091-6490

pii

1505914112

journal_volume

112

pub_type

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