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A Zea mays 39-kDa thylakoid transglutaminase catalyses the modification by polyamines of light-harvesting complex II in a light-dependent way

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Abstract

A transglutaminase (TGase; EC 2.3.2.13) activity, which shared many properties with the TGase activity of the Helianthus tuberosus chloroplast, was observed in the Zea mays L. chloroplast and in its fractions. This activity was found to be prevalent in thylakoids; bis-(glutamyl) spermidine and bis-(glutamyl) putrescine were the main polyamine conjugates formed. Light stimulated the endogenous thylakoid activity. Putrescine, spermidine and spermine were conjugated to the isolated light-harvesting complex of photosystem II (LHCII) with different degrees of efficiency, spermine being the polyamine most efficiently conjugated. A TGase with a light-sensitive activity was identified in the photosystem II-enriched fraction. Its partial purification on a sucrose gradient allowed the separation of a 39-kDa band, which was immunorecognised by two anti-TGase antibodies (Ab-3 and rat prostatic gland-TGase). Both a colorimetric and a radiometric assay for TGase activity, the former carried out in the presence of biotinylated cadaverine and the latter in the presence of polyamines labelled with radioactive isotopes and resulting in the isolation of glutamyl-polyamines, further confirmed that the thylakoid enzyme is indeed a calcium-dependent transglutaminase (Thyl-TGase). At variance with guinea pig liver and erythrocyte TGases, which are insensitive to light, the activity of the thylakoid transglutaminase is affected by light. Moreover, this enzyme, when tested with purified LHCII as substrate, catalysed the production of mono- and bis-glutamyl-polyamines in equal amounts, whereas the ‘animal’ enzymes produced mainly mono-derivatives. Herein, it is discussed whether this light sensitivity is due to the enzyme or the substrate.

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Abbreviations

bis-PU :

Bis-(γ-glutamyl) putrescine

bis-SD :

Bis-(γ-glutamyl) spermidine

bis-SM :

Bis-(γ-glutamyl) spermine

Er-TGase :

Erythrocyte transglutaminase

Gpl-TGase :

Guinea pig liver transglutaminase

LHCII :

Light-harvesting complex II

mono-PU :

Mono-(γ-glutamyl) putrescine

mono-SD :

Mono-(γ-glutamyl) spermidine

mono-SM :

Mono-(γ-glutamyl) spermine

PA :

Polyamine

PU :

Putrescine

SD :

Spermidine

SM :

Spermine

Rpg-TGase :

Rat prostatic gland transglutaminase

Thyl-TGase :

Thylakoid transglutaminase

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Acknowledgements

This work was supported by the University of Bologna (ex 60% and Funds for Selected Topics: ‘Segnali molecolari nel differenziamento cellulare’) and by”MIUR” Projects FIRB No. RBAU01KZ49 to Serafini-Fracassini; “MIUR” Projects FIRB No. RBAU01E3CX and FIRB No. RBNE01LACT to R.B. and F.V. We are very indebted to Prof. A. Serafini-Fracassini (Emeritus Professor of the University of St. Andrews, Scotland) for the restyling of the English manuscript. The authors are grateful to Prof. L. Moggi of the Dipartimento Ciamician, Gruppo Fotochimica, University of Bologna for the suggestions about the polyamine/chlorophyll interactions, to Henkel S.p.A. (Fino Mornasco, Como) for the kind gift of Deriphat C 160 KPC. We acknowledge the technical assistance of N. Mele for photographic and image assistance and of Dr. C. Pagnucco for HPLC analysis of some of the glutamyl-derivatives

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  1. L. Dondini

    Present address: Dipartimento di Colture Arboree, Università di Bologna, Via Fanin 46, 40126, Bologna, Italy

Authors and Affiliations

  1. Dipartimento di Biologia Evoluzionistica Sperimentale, Università di Bologna, Via Irnerio 42, 40126, Bologna, Italy

    M. Della Mea, A. Di Sandro, L. Dondini, S. Del Duca & D. Serafini-Fracassini

  2. Dipartimento Scientifico e Tecnologico, Università di Verona, Strada Le Grazie 15, 37134, Verona, Italy

    F. Vantini & R. Bassi

  3. Dipartimento di Biofisica e Biologia Molecolare, Università di Ferrara, Via L. Borsari 46, 44100, Ferrara, Italy

    C. Bergamini

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  1. M. Della Mea
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Correspondence to D. Serafini-Fracassini.

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Della Mea, M., Di Sandro, A., Dondini, L. et al. A Zea mays 39-kDa thylakoid transglutaminase catalyses the modification by polyamines of light-harvesting complex II in a light-dependent way. Planta 219, 754–764 (2004). https://doi.org/10.1007/s00425-004-1278-6

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