PDBsum entry 3pvb

Transferase

PDB id: 3pvb

Contents

Protein chains

345 a.a. *

160 a.a. *

Ligands

ANP

GOL

Metals

_MN ×2

Waters ×15

* Residue conservation analysis

PDB id:

3pvb

Name:

Transferase

Title:

Crystal structure of (73-244)ria:c holoenzyme of camp-dependent protein kinase

Structure:

Camp-dependent protein kinase catalytic subunit alpha. Chain: a. Fragment: unp residues 7-351. Synonym: pka c-alpha. Engineered: yes. Camp-dependent protein kinase type i-alpha regulatory subunit. Chain: b. Fragment: unp residues 85-244.

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: prkaca, pkaca. Expressed in: escherichia coli. Expression_system_taxid: 562. Bos taurus. Bovine,cow,domestic cattle,domestic cow. Organism_taxid: 9913.

Resolution:

3.30Å R-factor: 0.242 R-free: 0.290

Authors:

A.J.Boettcher,J.Wu,C.Kim,J.Yang,J.Bruystens,N.Cheung,J.K.Pennypacker, D.A.Blumenthal,A.P.Kornev,S.S.Taylor

Key ref:

A.J.Boettcher et al. (2011). Realizing the allosteric potential of the tetrameric protein kinase A RIα holoenzyme. Structure, 19, 265-276. PubMed id: 21300294

Date:

06-Dec-10 Release date: 02-Feb-11

Protein chain

P05132  (KAPCA_MOUSE) -  cAMP-dependent protein kinase catalytic subunit alpha from Mus musculus

Seq: 351 a.a.

Struc: 345 a.a.*

Protein chain

P00514  (KAP0_BOVIN) -  cAMP-dependent protein kinase type I-alpha regulatory subunit from Bos taurus

Seq: 380 a.a.

Struc: 160 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: Chain A: E.C.2.7.11.11 - cAMP-dependent protein kinase.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] (Bound ligand (Het Group name = ANP) matches with 81.25% similarity) + ADP + H(+)

L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] (Bound ligand (Het Group name = ANP) matches with 81.25% similarity) + ADP + H(+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

Structure 19:265-276 (2011)

PubMed id: 21300294

Realizing the allosteric potential of the tetrameric protein kinase A RIα holoenzyme.

A.J.Boettcher, J.Wu, C.Kim, J.Yang, J.Bruystens, N.Cheung, J.K.Pennypacker, D.A.Blumenthal, A.P.Kornev, S.S.Taylor.

ABSTRACT

PKA holoenzymes containing two catalytic (C) subunits and a regulatory (R) subunit dimer are activated cooperatively by cAMP. While cooperativity involves the two tandem cAMP binding domains in each R-subunit, additional cooperativity is associated with the tetramer. Of critical importance is the flexible linker in R that contains an inhibitor site (IS). While the IS becomes ordered in the R:C heterodimer, the overall conformation of the tetramer is mediated largely by the N-Linker that connects the D/D domain to the IS. To understand how the N-Linker contributes to assembly of tetrameric holoenzymes, we engineered a monomeric RIα that contains most of the N-Linker, RIα(73-244), and crystallized a holoenzyme complex. Part of the N-linker is now ordered by interactions with a symmetry-related dimer. This complex of two symmetry-related dimers forms a tetramer that reveals novel mechanisms for allosteric regulation and has many features associated with full-length holoenzyme. A model of the tetrameric holoenzyme, based on this structure, is consistent with previous small angle X-ray and neutron scattering data, and is validated with new SAXS data and with an RIα mutation localized to a novel interface unique to the tetramer.