39 research outputs found
Mechanism and Significance of Chlorophyll Breakdown
Chlorophyll breakdown is the most obvious sign of leaf senescence and fruit ripening. A multistep pathway has been elucidated in recent years that can be divided into two major parts. In the first phase, which commonly is active in higher plants, chlorophyll is converted via several photoreactive intermediates to a primary colorless breakdown product within the chloroplast. The second part of chlorophyll breakdown takes place in the cytosol and the vacuole. During this phase, the primary colorless intermediate is modified in largely species-specific reactions to a number of similar, yet structurally different, linear tetrapyrrolic products that finally are stored within the vacuole of senescing cells. To date, most of the biochemical reactions of the first phase of chlorophyll breakdown have been elucidated and genes have been identified. By contrast, mechanisms of catabolite transport and modification during the second phase are largely unknown. This review summarizes the current knowledge on the biochemical reactions involved in chlorophyll breakdown, with a special focus on the second-phase reactions and the fate of by-products that are released from chlorophyll during its breakdown
An evergreen mind and a heart for the colors of fall
With the finest biochemical and molecular approaches, convincing explorative strategies, and long-term vision, Stefan Hörtensteiner succeeded in elucidating the biochemical pathway responsible for chlorophyll degradation. After having contributed to the identification of key chlorophyll degradation products in the course of the past 25 years, he gradually identified and characterized most of the crucial players in the PAO/phyllobilin degradation pathway of chlorophyll. He was one of the brightest plant biochemists of his generation, and his work opened doors to a better understanding of plant senescence, tetrapyrrole homeostasis, and their complex regulation. He sadly passed away on 5 December 2020, aged 57
Non-specific activities of the major herbicide-resistance gene BAR
Bialaphos resistance (BAR) and phosphinothricin acetyltransferase (PAT) genes, which convey resistance to the broad-spectrum herbicide phosphinothricin (also known as glufosinate) via N-acetylation, have been globally used in basic plant research and genetically engineered crops1-4. Although early in vitro enzyme assays showed that recombinant BAR and PAT exhibit substrate preference toward phosphinothricin over the 20 proteinogenic amino acids1, indirect effects of BAR-containing transgenes in planta, including modified amino acid levels, have been seen but without the identification of their direct causes5,6. Combining metabolomics, plant genetics and biochemical approaches, we show that transgenic BAR indeed converts two plant endogenous amino acids, aminoadipate and tryptophan, to their respective N-acetylated products in several plant species. We report the crystal structures of BAR, and further delineate structural basis for its substrate selectivity and catalytic mechanism. Through structure-guided protein engineering, we generated several BAR variants that display significantly reduced non-specific activities compared with its wild-type counterpart in vivo. The transgenic expression of enzymes can result in unintended off-target metabolism arising from enzyme promiscuity. Understanding such phenomena at the mechanistic level can facilitate the design of maximally insulated systems featuring heterologously expressed enzymes.Searle Scholars Progra
A Role for TIC55 as a Hydroxylase of Phyllobilins, the Products of Chlorophyll Breakdown during Plant Senescence
Mechanism and significance of chlorophyll breakdown
Chlorophyll breakdown is the most obvious sign of leaf senescence and fruit ripening. A multistep pathway has been elucidated in recent years that can be divided into two major parts. In the first phase, which commonly is active in higher plants, chlorophyll is converted via several photoreactive intermediates to a primary colorless breakdown product within the chloroplast. The second part of chlorophyll breakdown takes place in the cytosol and the vacuole. During this phase, the primary colorless intermediate is modified in largely species-specific reactions to a number of similar, yet structurally different, linear tetrapyrrolic products that finally are stored within the vacuole of senescing cells. To date, most of the biochemical reactions of the first phase of chlorophyll breakdown have been elucidated and genes have been identified. By contrast, mechanisms of catabolite transport and modification during the second phase are largely unknown. This review summarizes the current knowledge on the biochemical reactions involved in chlorophyll breakdown, with a special focus on the second-phase reactions and the fate of by-products that are released from chlorophyll during its breakdown
Characterization of the pheophorbide a oxygenase/phyllobilin pathway of chlorophyll breakdown in grasses
MAIN CONCLUSION
Although the PAO/phyllobilin pathway of chlorophyll breakdown is active in grass leaf senescence, the abundance of phyllobilins is far below the amount of degraded chlorophyll. The yellowing of fully developed leaves is the most prominent visual symptom of plant senescence. Thereby, chlorophyll is degraded via the so-called pheophorbide a oxygenase (PAO)/phyllobilin pathway to a species-specific set of phyllobilins, linear tetrapyrrolic products of chlorophyll breakdown. Here, we investigated the diversity and abundance of phyllobilins in cereal and forage crops, i.e. barley, rice, ryegrass, sorghum and wheat, using liquid chromatography-mass spectrometry. A total of thirteen phyllobilins were identified, among them four novel, not yet described ones, pointing to a rather high diversity of phyllobilin-modifying activities present in the Gramineae. Along with these phyllobilins, barley orthologs of known Arabidopsis thaliana chlorophyll catabolic enzymes were demonstrated to localize in the chloroplast, and two of them, i.e. PAO and pheophytin pheophorbide hydrolase, complemented respective Arabidopsis mutants. These data confirm functionality of the PAO/phyllobilin pathway in grasses. Interestingly, when comparing phyllobilin abundance with amounts of degraded chlorophyll in senescent leaves, in most analyzed grass species only minor fractions of chlorophyll were recovered as phyllobilins, opposite to A. thaliana where phyllobilin quantities match degraded chlorophyll rather well. These data show that, despite the presence and activity of the PAO/phyllobilin pathway in barley (and other cereals), phyllobilins do not accumulate stoichiometrically, implying possible degradation of chlorophyll beyond the phyllobilin level
Hydroxymethylated phyllobilins: a puzzling new feature of the dioxobilin branch of chlorophyll breakdown
Colorless nonfluorescent chlorophyll (Chl) catabolites (NCCs) are formyloxobilin-type phyllobilins, which are considered the typical products of Chl breakdown in senescent leaves. However, in degreened leaves of some plants, dioxobilin-type Chl catabolites (DCCs) predominate, which lack the formyl group of the NCCs, and which arise from Chl catabolites by oxidative removal of the formyl group by a P450 enzyme. Here a structural investigation of the DCCs in the methylesterase16 mutant of Arabidopsis thaliana is reported. Eight new DCCs were identified and characterized structurally. Strikingly, three of these DCCs carry stereospecifically added hydroxymethyl groups, and represent bilin-type linear tetrapyrroles with an unprecedented modification. Indeed, DCCs show a remarkable structural parallel, otherwise, to the bilins from heme breakdown