PDBsum entry 1cx4

Signaling protein

PDB id: 1cx4

Contents

Protein chain

275 a.a. *

Ligands

CMP ×2

Waters ×192

* Residue conservation analysis

References listed in PDB file

Key reference

• Title: Molecular basis for regulatory subunit diversity in camp-Dependent protein kinase: crystal structure of the type ii beta regulatory subunit.
• Authors: T.C.Diller, Madhusudan, N.H.Xuong, S.S.Taylor.
• Ref.: Structure, 2001, 9, 73-82. [DOI no: 10.1016/S0969-2126(00)00556-6]
• PubMed id: 11342137

Abstract

BACKGROUND: Cyclic AMP binding domains possess common structural features yet are diversely coupled to different signaling modules. Each cAMP binding domain receives and transmits a cAMP signal; however, the signaling networks differ even within the same family of regulatory proteins as evidenced by the long-standing biochemical and physiological differences between type I and type II regulatory subunits of cAMP-dependent protein kinase. RESULTS: We report the first type II regulatory subunit crystal structure, which we determined to 2.45 A resolution and refined to an R factor of 0.176 with a free R factor of 0.198. This new structure of the type II beta regulatory subunit of cAMP-dependent protein kinase demonstrates that the relative orientations of the two tandem cAMP binding domains are very different in the type II beta as compared to the type I alpha regulatory subunit. Each structural unit for binding cAMP contains the highly conserved phosphate binding cassette that can be considered the "signature" motif of cAMP binding domains. This motif is coupled to nonconserved regions that link the cAMP signal to diverse structural and functional modules. CONCLUSIONS: Both the diversity and similarity of cAMP binding sites are demonstrated by this new type II regulatory subunit structure. The structure represents an intramolecular paradigm for the cooperative triad that links two cAMP binding sites through a domain interface to the catalytic subunit of cAMP-dependent protein kinase. The domain interface surface is created by the binding of only one cAMP molecule and is enabled by amino acid sequence variability within the peptide chain that tethers the two domains together.

Figure 4.
Figure 4. Hydrogen Bonds Surrounding cAMPThe network of conserved and nonconserved hydrogen bonds surrounding the two cAMP binding sites of RIIb.(a) Binding site A.(b) Binding site B. Cyclic AMP molecules are rendered as balls and sticks with gray for carbon, blue for nitrogen, red for oxygen, and purple for the phosphorous atom. Hydrogen bonding is represented by dashed lines. Water oxygen atoms are illustrated as ovals with gradient shading and the uniquely identifying three-digit numeral assigned by the PDB file 1cx4. Tan-shaded boxes signify secondary structural elements identified in Figure 3; the labels for these boxes are either red for b strands or green for a helices. The regions encompassed by yellow lie within the conserved phosphate binding cassette. Amino acid residues with more than one hydrogen-bonding atom are boxed and labeled in either blue for invariant residues or black for nonconserved residues

The above figure is reprinted by permission from Cell Press: Structure (2001, 9, 73-82) copyright 2001.

Secondary reference #1

• Title: Type ii beta regulatory subunit of camp-Dependent protein kinase: purification strategies to optimize crystallization.
• Authors: T.C.Diller, N.H.Xuong, S.S.Taylor.
• Ref.: Protein Expr Purif, 2000, 20, 357-364. [DOI no: 10.1006/prep.2000.1312]
• PubMed id: 11087674