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Structural insight into nucleotide recognition by human death-associated protein kinase.

McNamara LK, Watterson DM, Brunzelle JS - Acta Crystallogr. D Biol. Crystallogr. (2009)

Bottom Line: Death-associated protein kinase (DAPK) is a member of the Ca(2+)/calmodulin-regulated family of serine/threonine protein kinases.The structure of DAPK-ADP-Mg(2+) was compared with a newly determined DAPK-AMP-PNP-Mg(2+) structure and the previously determined apo DAPK structure (PDB code 1jks).The comparison shows that nucleotide-induced changes are localized to the glycine-rich loop region of DAPK.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Drug Discovery and Chemical Biology, Northwestern University, Chicago, Illinois 60611, USA.

ABSTRACT
Death-associated protein kinase (DAPK) is a member of the Ca(2+)/calmodulin-regulated family of serine/threonine protein kinases. The role of the kinase activity of DAPK in eukaryotic cell apoptosis and the ability of bioavailable DAPK inhibitors to rescue neuronal death after brain injury have made it a drug-discovery target for neurodegenerative disorders. In order to understand the recognition of nucleotides by DAPK and to gain insight into DAPK catalysis, the crystal structure of human DAPK was solved in complex with ADP and Mg(2+) at 1.85 A resolution. ADP is a product of the kinase reaction and product release is considered to be the rate-limiting step of protein kinase catalytic cycles. The structure of DAPK-ADP-Mg(2+) was compared with a newly determined DAPK-AMP-PNP-Mg(2+) structure and the previously determined apo DAPK structure (PDB code 1jks). The comparison shows that nucleotide-induced changes are localized to the glycine-rich loop region of DAPK.

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The structure of DAPK–ADP–Mg2+. (a) A representation of the overall structure of DAPK–ADP–Mg2+. (b) An F                  o − F                  c density map calculated with ADP and Mg2+ omitted from the model is shown contoured at +3σ within the context of the surrounding protein residues.
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fig1: The structure of DAPK–ADP–Mg2+. (a) A representation of the overall structure of DAPK–ADP–Mg2+. (b) An F o − F c density map calculated with ADP and Mg2+ omitted from the model is shown contoured at +3σ within the context of the surrounding protein residues.

Mentions: The structure of DAPK–ADP–Mg2+ (Fig. 1 ▶ a) shows that the structure has the canonical kinase fold consisting of a mainly N-terminal β-sheet and a larger α-helical C-terminal domain. The F o − F c OMIT map shows density corresponding to the ADP molecule between the N-terminal and C-­terminal domains (Fig. 1 ▶ b). Density for the adenine ring, the ribose ring, phosphates and one magnesium ion is clearly identifiable.


Structural insight into nucleotide recognition by human death-associated protein kinase.

McNamara LK, Watterson DM, Brunzelle JS - Acta Crystallogr. D Biol. Crystallogr. (2009)

The structure of DAPK–ADP–Mg2+. (a) A representation of the overall structure of DAPK–ADP–Mg2+. (b) An F                  o − F                  c density map calculated with ADP and Mg2+ omitted from the model is shown contoured at +3σ within the context of the surrounding protein residues.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651756&req=5

fig1: The structure of DAPK–ADP–Mg2+. (a) A representation of the overall structure of DAPK–ADP–Mg2+. (b) An F o − F c density map calculated with ADP and Mg2+ omitted from the model is shown contoured at +3σ within the context of the surrounding protein residues.
Mentions: The structure of DAPK–ADP–Mg2+ (Fig. 1 ▶ a) shows that the structure has the canonical kinase fold consisting of a mainly N-terminal β-sheet and a larger α-helical C-terminal domain. The F o − F c OMIT map shows density corresponding to the ADP molecule between the N-terminal and C-­terminal domains (Fig. 1 ▶ b). Density for the adenine ring, the ribose ring, phosphates and one magnesium ion is clearly identifiable.

Bottom Line: Death-associated protein kinase (DAPK) is a member of the Ca(2+)/calmodulin-regulated family of serine/threonine protein kinases.The structure of DAPK-ADP-Mg(2+) was compared with a newly determined DAPK-AMP-PNP-Mg(2+) structure and the previously determined apo DAPK structure (PDB code 1jks).The comparison shows that nucleotide-induced changes are localized to the glycine-rich loop region of DAPK.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Drug Discovery and Chemical Biology, Northwestern University, Chicago, Illinois 60611, USA.

ABSTRACT
Death-associated protein kinase (DAPK) is a member of the Ca(2+)/calmodulin-regulated family of serine/threonine protein kinases. The role of the kinase activity of DAPK in eukaryotic cell apoptosis and the ability of bioavailable DAPK inhibitors to rescue neuronal death after brain injury have made it a drug-discovery target for neurodegenerative disorders. In order to understand the recognition of nucleotides by DAPK and to gain insight into DAPK catalysis, the crystal structure of human DAPK was solved in complex with ADP and Mg(2+) at 1.85 A resolution. ADP is a product of the kinase reaction and product release is considered to be the rate-limiting step of protein kinase catalytic cycles. The structure of DAPK-ADP-Mg(2+) was compared with a newly determined DAPK-AMP-PNP-Mg(2+) structure and the previously determined apo DAPK structure (PDB code 1jks). The comparison shows that nucleotide-induced changes are localized to the glycine-rich loop region of DAPK.

Show MeSH
Related in: MedlinePlus