Annotations for this protein have been verified by the authors of the corresponding papers


DP00534: Calvin cycle protein CP12-2, chloroplasticFASTA viewXML view

General information
DisProt:DP00534
Name:Calvin cycle protein CP12-2, chloroplastic
Synonym(s):CP122_ARATH
CP12 domain-containing protein 2
Chloroplast protein 12-2
CP12 protein-like protein
First appeared in release:Release 3.7 (01/28/2008)
UniProt:Q9LZP9
UniGene: 
SwissProt: CP122_ARATH
TrEMBL:  
NCBI (GI): 75155154
Source organism:Arabidopsis thaliana (Mouse-ear cress)
Sequence length:131
Percent disordered:60%
Homologues: 


Native sequence

        10         20         30         40         50         60
         |          |          |          |          |          |
MATIATGLNI ATQRVFVTSE NRPVCLAGPV HLNNSWNLGS RTTNRMMKLQ PIKAAPEGGI - 60
SDVVEKSIKE AQETCAGDPV SGECVAAWDE VEELSAAASH ARDKKKADGS DPLEEYCKDN - 120
PETNECRTYD N



Functional narrative    

Chloroplastic Calvin cycle protein CP12-2 contains a transit peptide at aa M1-K53; the mature protein comprises A54-N131.

CP12 is a protein of 8.7 kDa that contributes to regulation of the Calvin cycle by acting as a scaffold element in the formation of a supramolecular complex with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) in photosynthetic organisms. CD and NMR studies of recombinant CP12 (isoform 2) of Arabidopsis thaliana show that CP12-2 is mostly unstructured.

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Function: Acts as a linker essential in the assembly of a core complex of PRK/GAPDH. Coordinates the reversible inactivation of chloroplast enzymes GAPDH and PRK during darkness in photosynthetic tissues.

Subunit structure: Monomer. Component of a complex that contains two dimers of PRK, two tetramers of GAPDH and four CP12. CP12 associates with GAPDH, causing its conformation to change. This GAPDH/(CP12)2 complex binds PRK to form a half-complex (one unit). This unit probably dimerizes due partially to interactions between the enzymes of each unit.

Subcellular location: Plastid › chloroplast
Tissue specificity: Mostly expressed in cotyledons, leaves and flower stalks, and, to a lower extent, in flowers and stems. Barely detectable in roots and siliques.
Developmental stage: In flowers, expressed in the sepals and the style of flowers. In siliques, present at the base and tip.
Induction: Induced by light. Repressed by darkness, cold, anaerobic treatment, heat and sucrose.

Post-translational modification: Contains two disulfide bonds; Changes conformation depending on redox conditions. Only the oxidized protein, with two disulfide bonds, is active in complex formation. The C-terminal disulfide is involved in the interaction with GAPDH and the N-terminal disulfide mediates the binding of PRK with this binary complex.

Sequence similarities: Belongs to the CP12 family.
Redox potential: E0 are -326 mV and -352 mV for the disulfide bonds at pH 7.9.
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Region 2: 54-109 Region 1: 54-131 Region 3: 110-131

Map of ordered and disordered regions







Note: 'Mouse' over a region to see the start and stop residues. Click on a region to see detailed information.


Region 1
Type:Disordered
Name:CP12-2 (mature protein)
Location:54 - 131
Length:78
Region sequence:

AAPEGGISDVVEKSIKEAQETCAGDPVSGECVAAWDEVEELSAAASHARDKKKADGSDPL
EEYCKDNPETNECRTYDN

Modification type: Complex
Native
PDB:  
Structural/functional type: Function arises from the disordered state
Functional classes: Molecular assembly
Functional subclasses: Protein-protein binding
Detection methods:
  1. Nuclear magnetic resonance (NMR) (293 K; pH: 7; potassium phosphate 25 mM; CP12-2 1 mM; DTT (oxidizing agent) 20 mM; sodium azide (0.05 w/v); 95% H2O/5% D2O)

  2. Dynamic light scattering (pH: 7.5; CP12-2 (singly, or in pre-formed binary or ternary complex); EDTA 1 mM; KCl 150 mM; NAD (for complexes) 0.2 mM; potassium phosphate 25 mM)
References:
  1. Marri L, Trost P, Pupillo P, Sparla F. "Reconstitution and properties of the recombinant glyceraldehyde-3-phosphate dehydrogenase/CP12/phosphoribulokinase supramolecular complex of Arabidopsis." Plant Physiol.. 2005; 139(3): 1433-43. PubMed: 16258009

  2. Marri L, Trost P, Trivelli X, Gonnelli L, Pupillo P, Sparla F. "Spontaneous assembly of photosynthetic supramolecular complexes as mediated by the intrinsically unstructured protein CP12." J Biol Chem. 2007. PubMed: 17947231
Comments:
 



Region 2
Type:Disordered
Name:N-terminal region
Location:54 - 109
Length:56
Region sequence:

AAPEGGISDVVEKSIKEAQETCAGDPVSGECVAAWDEVEELSAAASHARDKKKADG

Modification type: Complex
Engineered
PDB: 2LJ9:A, 3QV1:G, 3RVD:I
Structural/functional type: Function arises from the disordered state
Functional classes: Molecular assembly
Functional subclasses: Protein-protein binding
Detection methods:
  1. Nuclear magnetic resonance (NMR) (293 K; pH: 7; 95% H2O/5% D2O (or 100% D2O); CP12, aa A54-N131 (PDB 2LJ9) 1 mM; potassium phosphate 25 mM; sodium azide (0.05 w/v); TSP (0.1-0.2 mM) 0.2 mM)

  2. X-ray crystallography (277 K; pH: 6.5; CP12, in complex with GAPDH (PDB 3QV1 [co-crystallization experiments]); MES 0.1 M; NaCl 0.6 M; NAD 1 mM; PEG 4K 15 %)

  3. X-ray crystallography (293 K; ammonium sulphate 2.4 M; CP12, in complex with GAPDH (PDB 3RVD [soaking experiments]) 3.125 mg/mL; NAD 1 mM; sodium citrate 0.1 M)

  4. Circular dichroism (CD) spectroscopy, far-UV (293 K; pH: 7; citrate 15 mM; CP12 100 uM)
References:
  1. Fermani S, Trivelli X, Sparla F, Thumiger A, Calvaresi M, Marri L, Falini G, Zerbetto F, Trost P. "Conformational selection and folding-upon-binding of intrinsically disordered protein CP12 regulate photosynthetic enzymes assembly." J. Biol. Chem.. 2012; 287(25): 21372-83. PubMed: 22514274
Comments:
According to Fermani et al (2012), oxidized CP12-2 (mature form, two disulfide bonds) is disordered from A54-G109 and from R127-N131. The segment S110-C126 is more ordered and includes an alpha-helix and a loop linked together by a disulfide bridge. The whole C-terminal region from S110-N131 shows disorder-to-order transition upon binding to GAPDH.


Region 3
Type:Disordered w/ Residual Structure
Name:C-terminal region
Location:110 - 131
Length:22
Region sequence:

SDPLEEYCKDNPETNECRTYDN

Modification type: Complex
Engineered
PDB: 2LJ9:A, 3QV1:G, 3RVD:I
Structural/functional type: Function arises via a disorder to order transition
Functional classes: Molecular assembly
Functional subclasses: Protein-protein binding
Detection methods:
  1. Nuclear magnetic resonance (NMR) (293 K; pH: 7; 95% H2O/5% D2O (or 100% D2O); CP12, aa A54-N131 (PDB 2LJ9) 1 ; potassium phosphate 25 mM; sodium azide (0.05 w/v); TSP (0.1-0.2 mM) 0.2 mM)

  2. X-ray crystallography (277 K; pH: 6.5; CP12, in complex with GAPDH (PDB 3QV1 [co-crystallization experiments]); MES 0.1 M; NaCl 0.6 M; NAD 1 mM; PEG 4K 15 %)

  3. X-ray crystallography (293 K; ammonium sulphate 2.4 M; CP12, in complex with GAPDH 3.125 mg/mL; NAD 1 mM; sodium citrate 0.1 M)

  4. Circular dichroism (CD) spectroscopy, far-UV (293 K; citrate 15 mM; CP12 100 uM)
References:
  1. Fermani S, Trivelli X, Sparla F, Thumiger A, Calvaresi M, Marri L, Falini G, Zerbetto F, Trost P. "Conformational selection and folding-upon-binding of intrinsically disordered protein CP12 regulate photosynthetic enzymes assembly." J. Biol. Chem.. 2012; 287(25): 21372-83. PubMed: 22514274
Comments:
According to Fermani et al (2012), oxidized CP12-2 (mature form, two disulfide bonds) is disordered from A54-G109 and from R127-N131. The segment S110-C126 is more ordered and includes an alpha-helix and a loop linked together by a disulfide bridge. The whole C-terminal region from S110-N131 shows disorder-to-order transition upon binding to GAPDH.


References

  1. Fermani S, Trivelli X, Sparla F, Thumiger A, Calvaresi M, Marri L, Falini G, Zerbetto F, Trost P. "Conformational selection and folding-upon-binding of intrinsically disordered protein CP12 regulate photosynthetic enzymes assembly." J. Biol. Chem.. 2012; 287(25): 21372-83. PubMed: 22514274

  2. Kleffmann T, Russenberger D, von Zychlinski A, Christopher W, Sjölander K, Gruissem W, Baginsky S. "The Arabidopsis thaliana chloroplast proteome reveals pathway abundance and novel protein functions." Curr. Biol.. 2004; 14(5): 354-62. PubMed: 15028209

  3. Marri L, Pesaresi A, Valerio C, Lamba D, Pupillo P, Trost P, Sparla F. "In vitro characterization of Arabidopsis CP12 isoforms reveals common biochemical and molecular properties." J. Plant Physiol.. 2010; 167(12): 939-50. PubMed: 20399532

  4. Marri L, Sparla F, Pupillo P, Trost P. "Co-ordinated gene expression of photosynthetic glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, and CP12 in Arabidopsis thaliana." J. Exp. Bot.. 2004; 56(409): 73-80. PubMed: 15533878

  5. Marri L, Trost P, Pupillo P, Sparla F. "Reconstitution and properties of the recombinant glyceraldehyde-3-phosphate dehydrogenase/CP12/phosphoribulokinase supramolecular complex of Arabidopsis." Plant Physiol.. 2005; 139(3): 1433-43. PubMed: 16258009

  6. Marri L, Trost P, Trivelli X, Gonnelli L, Pupillo P, Sparla F. "Spontaneous assembly of photosynthetic supramolecular complexes as mediated by the intrinsically unstructured protein CP12." J Biol Chem. 2007. PubMed: 17947231

  7. Olinares PD, Ponnala L, van Wijk KJ. "Megadalton complexes in the chloroplast stroma of Arabidopsis thaliana characterized by size exclusion chromatography, mass spectrometry, and hierarchical clustering." Mol. Cell Proteomics. 2010; 9(7): 1594-615. PubMed: 20423899

  8. Singh P, Kaloudas D, Raines CA. "Expression analysis of the Arabidopsis CP12 gene family suggests novel roles for these proteins in roots and floral tissues." J. Exp. Bot.. 2008; 59(14): 3975-85. PubMed: 18974062

  9. Zybailov B, Rutschow H, Friso G, Rudella A, Emanuelsson O, Sun Q, van Wijk KJ. "Sorting signals, N-terminal modifications and abundance of the chloroplast proteome." PLoS One. 2008; 3(4): e1994. PubMed: 18431481



Comments


AV sent 9/10/1/2012 (PMID 22514274)
Verified 10-5-2012


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