A Zea mays 39-kDa thylakoid transglutaminase catalyses the modification by polyamines of light-harvesting complex II in a light-dependent way.
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.
Della Mea
M
M
Dipartimento di Biologia Evoluzionistica Sperimentale, Università di Bologna, Via Irnerio 42, 40126, Bologna, Italy.
Di Sandro
A
A
Dondini
L
L
Del Duca
S
S
Vantini
F
F
Bergamini
C
C
Bassi
R
R
Serafini-Fracassini
D
D
D000818:Animals; D002103:Cadaverine / Q000378:metabolism; D008027:Light; D008297:Male; D045332:Photosystem II Protein Complex / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism*; D011467:Prostate / Q000201:enzymology; D051381:Rats; D020524:Thylakoids / Q000201:enzymology*; D011503:Transglutaminases / Q000378:metabolism*; D003313:Zea mays / Q000201:enzymology*
D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D011073:Polyamines; D011503:Transglutaminases; D002103:Cadaverine