Isolation and Compositional analysis of a CP12-associated complex of Calvin Cycle enzymes from Nicotiana tabacum

Lancaster Environment Centre

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https://doi.org/10.2174/092986611795222740
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Keywords

Calvin Cycle, CP12, Glyceraldehyde-3-phosphate dehydrogenase, Phosphoribulokinase, Protein-protein interactions, Redox regulation

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Isolation and Compositional analysis of a CP12-associated complex of Calvin Cycle enzymes from Nicotiana tabacum. / Carmo-Silva, A. Elizabete; Marri, Lucia; Sparla, Francesca et al.
In: Protein and Peptide Letters, Vol. 18, No. 6, 01.06.2011, p. 618-624.

Research output: Contribution to Journal/Magazine › Journal article › peer-review

Bibtex

@article{b0ca223eb5714b9f9a3ece6c2ac37706,

title = "Isolation and Compositional analysis of a CP12-associated complex of Calvin Cycle enzymes from Nicotiana tabacum",

abstract = "Two Calvin Cycle enzymes, NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) form a multiprotein complex with CP12, a small intrinsically-unstructured protein. Under oxidizing conditions, association with CP12 confers redox-sensitivity to the otherwise redox-insensitive A isoform of GAPDH (GapA) and provides an additional level of down-regulation to the redox-regulated PRK. To determine if CP12- mediated regulation is specific for GAPDH and PRK in vivo, a high molecular weight complex containing CP12 was isolated from tobacco chloroplasts and leaves and its protein composition was characterized. Gel electrophoresis and immunoblot analyses after separation of stromal proteins by size fractionation verified that the GAPDH (both isoforms) and PRK co-migrated with CP12 in dark- but not light-adapted chloroplasts. Nano-liquid-chromatography-mass-spectrometry of the isolated complex identified only CP12, GAPDH and PRK. Since nearly all of the CP12 from darkened chloroplasts migrates with GADPH and PRK as a high molecular mass species, these data indicate that the tight association of tobacco CP12 with GAPDH and PRK is specific and involves no other Calvin Cycle enzymes.",

keywords = "Calvin Cycle, CP12, Glyceraldehyde-3-phosphate dehydrogenase, Phosphoribulokinase, Protein-protein interactions, Redox regulation",

author = "Carmo-Silva, {A. Elizabete} and Lucia Marri and Francesca Sparla and Salvucci, {Michael E.}",

year = "2011",

month = jun,

day = "1",

doi = "10.2174/092986611795222740",

language = "English",

volume = "18",

pages = "618--624",

journal = "Protein and Peptide Letters",

issn = "0929-8665",

publisher = "Bentham Science Publishers B.V.",

number = "6",

}

RIS

TY - JOUR

T1 - Isolation and Compositional analysis of a CP12-associated complex of Calvin Cycle enzymes from Nicotiana tabacum

AU - Carmo-Silva, A. Elizabete

AU - Marri, Lucia

AU - Sparla, Francesca

AU - Salvucci, Michael E.

PY - 2011/6/1

Y1 - 2011/6/1

N2 - Two Calvin Cycle enzymes, NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) form a multiprotein complex with CP12, a small intrinsically-unstructured protein. Under oxidizing conditions, association with CP12 confers redox-sensitivity to the otherwise redox-insensitive A isoform of GAPDH (GapA) and provides an additional level of down-regulation to the redox-regulated PRK. To determine if CP12- mediated regulation is specific for GAPDH and PRK in vivo, a high molecular weight complex containing CP12 was isolated from tobacco chloroplasts and leaves and its protein composition was characterized. Gel electrophoresis and immunoblot analyses after separation of stromal proteins by size fractionation verified that the GAPDH (both isoforms) and PRK co-migrated with CP12 in dark- but not light-adapted chloroplasts. Nano-liquid-chromatography-mass-spectrometry of the isolated complex identified only CP12, GAPDH and PRK. Since nearly all of the CP12 from darkened chloroplasts migrates with GADPH and PRK as a high molecular mass species, these data indicate that the tight association of tobacco CP12 with GAPDH and PRK is specific and involves no other Calvin Cycle enzymes.

AB - Two Calvin Cycle enzymes, NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) form a multiprotein complex with CP12, a small intrinsically-unstructured protein. Under oxidizing conditions, association with CP12 confers redox-sensitivity to the otherwise redox-insensitive A isoform of GAPDH (GapA) and provides an additional level of down-regulation to the redox-regulated PRK. To determine if CP12- mediated regulation is specific for GAPDH and PRK in vivo, a high molecular weight complex containing CP12 was isolated from tobacco chloroplasts and leaves and its protein composition was characterized. Gel electrophoresis and immunoblot analyses after separation of stromal proteins by size fractionation verified that the GAPDH (both isoforms) and PRK co-migrated with CP12 in dark- but not light-adapted chloroplasts. Nano-liquid-chromatography-mass-spectrometry of the isolated complex identified only CP12, GAPDH and PRK. Since nearly all of the CP12 from darkened chloroplasts migrates with GADPH and PRK as a high molecular mass species, these data indicate that the tight association of tobacco CP12 with GAPDH and PRK is specific and involves no other Calvin Cycle enzymes.

KW - Calvin Cycle

KW - CP12

KW - Glyceraldehyde-3-phosphate dehydrogenase

KW - Phosphoribulokinase

KW - Protein-protein interactions

KW - Redox regulation

U2 - 10.2174/092986611795222740

DO - 10.2174/092986611795222740

M3 - Journal article

C2 - 21271977

AN - SCOPUS:79953331751

VL - 18

SP - 618

EP - 624

JO - Protein and Peptide Letters

JF - Protein and Peptide Letters

SN - 0929-8665

IS - 6

ER -

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