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Identification of chlorophyll-a/b proteins as substrates of transglutaminase activity in isolated chloroplasts of Helianthus tuberosus L.

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Abstract

Endogenous substrates of transglutaminase (TGase; EC 2.3.2.13) have been identified in choloroplasts of Helianthus tuberosus leaves. The activity of TGase is Ca2+- and light-stimulated and catalyzes the incorporation of polyamines into thylakoid and stromal proteins. These proteins were separated by two-dimensional gel electrophoresis (first dimension: Deriphat-PAGE; second dimension: SDS-urea-PAGE) and Western-blotted. The thylakoid proteins were recognized by polyclonal antibodies as apoproteins of the chlorophyll-a/b antenna complex (LHCII, CP24, CP26 and CP29); a stromal protein was recognized by antibodies as the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. A possible localization of the acyl donor site for CP26 is proposed. A comparative analysis of polyamine incorporation into trichloroacetic-acid-precipitable material indicated that spermidine was a preferential acyl-acceptor substrate with respect to putrescine, even though the above-reported substrates are the same. The nature of the substrates, together with the light stimulation, support the working hypothesis of a possible role of TGase in regulating the light-harvesting function.

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Abbreviations

CP:

chlorophyll protein

LHC:

light-harvesting complex

Mr :

relative molecular mass

PA:

polyamine

PU:

putrescine

Rubisco:

ribulose-1,5-bisphosphate carboxylase/oxygenase

SD:

spermidine

TCA:

trichloroacetic acid

TGase:

transglutaminase

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Authors and Affiliations

  1. Dipartimento di Biologia Evoluzionistica Sperimentale, Università di Bologna, via Irnerio 42, I-40126, Bologna, Italy

    S. Del Duca & D. Serafmi-Fracassini

  2. Dipartimento di Biologia, Università di Padova, via Trieste 75, I-35100, Padova, Italy

    V. Tidu

  3. Dipartimento di Biotecnologie Vegetali, Università di Verona, strada Le Grazie 5, I-37100, Verona, Italy

    R. Bassi

  4. Dipartimento di Biochimica e Biofisica, Università di Napoli, Via Costantinopoli 16, I-80138, Napoli, Italy

    C. Esposito

Authors
  1. S. Del Duca
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  2. V. Tidu
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  3. R. Bassi
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  4. C. Esposito
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  5. D. Serafmi-Fracassini
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Additional information

We acknowledge the financial support provided by CNR contribution No. 91.00539.CTO 4 to D. Serafini-Fracassini and MAF grant No. 4.7240.90 to R. Bassi. We thank Prof. C. Bergamini (Istituto di Biochimica, Università di Ferrara, Ferrara, Italy) for the kind gift of antibodies against human erythrocyte TGase.

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Del Duca, S., Tidu, V., Bassi, R. et al. Identification of chlorophyll-a/b proteins as substrates of transglutaminase activity in isolated chloroplasts of Helianthus tuberosus L.. Planta 193, 283–289 (1994). https://doi.org/10.1007/BF00192542

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  • DOI: https://doi.org/10.1007/BF00192542

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