Protochlorophyllide photoreduction

Chlorophylls play a fundamental role in the absorption of light energy and its conversion into chemical energy for use by all organisms living on the Earth. The formation of these compounds occurs by a complex series of react ions carried out throughout the lifetime of the plant. One step of this rea ction series, the conversion of protochlorophyllide into chlorophyllide, is unique in its requirement for light. The reduction of protochlorophyllide to chlorophyllide is catalyzed by the nuclear-encoded enzyme NADPH: protoch lorophyllide oxidoreductase (POR). Recently, genes encoding two different P OR proteins, designated PORa and PORb, have been identified in vascular pla nts. Despite a high degree of amino acid sequence similarity, the roles pla yed by these different gene products appear to be quite distinct during pho tomorphogenesis. While PORb is present and active throughout the life of th e plant, PORa appears to be present and functional only in the first few ho urs of greening following the onset of illumination of etiolated (or dark-a dapted) plants. Consistent with this, analysis of gene expression patterns showed that the transcription of the PORa and PORb encoding genes are contr olled differently by light and plant developmental stage. PORa transcriptio n is negatively photoregulated by light. Phytochrome, mainly Phytochrome a, is responsible for light control of the gene transcription. Import of PORa but not PORb into plastids also appears to be differentially regulated, wi th pPORa translocation across the chloroplast envelope being controlled by substrate (protochlorophyllide) availability. In etiolated plants, POR form s large aggregates located in tube-like structures termed, 'prolamellar bod ies'. Some amount of POR was detected in prothylakoids. In light-adapted va scular plants, green algae and cyanobacteria, POR and protochlorophyllide w ere detected in chloroplast envelope and stroma membranes. Photoactive enzy me complexes are identified mainly in stroma membranes near polyribosomes. The unique feature of POR is that it is a photo-enzyme whose catalytic acti vity depends on light. It also requires NADPH and undergoes conformational changes near Cys groups in the course of catalytic activity. The first step of the reaction is photo-induced electron transfer to protoc hlorophyllide that leads to the formation of a nonfluorescent ion-radical. This step is followed by hydrogen transfer from NADPH. Degradation of PORa is specifically controlled and performed by the nuclear encoded proteases. The mechanism of the light activation of these proteases expression is unkn own, but they appear only after onset of illumination. Their activity also depends on the susceptibility of POR after its conformational change follow ed by protochlorophyllide photoreduction. Analysis of chlorophyll biosynthe sis in chloroplasts of light-adapted plants revealed a new mechanism of pro tochlorophyllide photo-reduction. The reaction differs from that previously observed in etiolated plants by the initial photoactive protochlorophyllid e form, intermediate steps and the final product. The rates of chlorophyll accumulation through these two reactions also differ. The same reaction was also detected in etiolated plants in parallel to the main reaction of prot ochlorophyllide photoreduction. Study of different photosynthetic organisms reveals the presence of POR in all of them including higher plants, green algae and cyanobacteria, which possess for light-independent chlorophyll sy nthesis, except for photosynthetic bacteria. Among those, PORa is present o nly in angiosperms, which lost the ability to synthesize chlorophyll in the dark. Sequence similarity reveals the evolutionary origin of FOR from shor t-chain alcohol dehydrogenases.