Biology subjects

Proctor, M. S.

Publications and source records attributed to Proctor, M. S..

4 recordsLinked to original sources

bioRxiv2026

Structure of chlorophyll synthase in complex with the LHC-like protein HliD

Chlorophyll is the central cofactor in oxygenic photosynthesis, responsible for both light capture in antenna complexes and light-driven charge separation in photosystems. The tetraprenyl tail of chlorophyll is attached to the chlorophyllide macrocycle by the transmembrane enzyme chlorophyll synthase (ChlG). In the cyanobacterium Synechocystis sp. PCC 6803, ChlG forms a complex with the LHC-like high-light-inducible protein HliD. To understand the substrate specificity and catalytic mechanism of ChlG, and how it is photoprotected by HliD, we determined the structure of a ChlG2HliD2 complex in both substrate-free and geranylgeranyl pyrophosphate (GGPP)-bound states using cryogenic electron microscopy (cryo-EM). A homodimer of HliD, which binds four chlorophylls and two zeaxanthin molecules in a quenched state, is flanked by two ChlG monomers. AlphaFold modelling placed chlorophyllide adjacent to the structurally resolved GGPP bound to the active site of ChlG. Cryo-EM data, site-directed mutagenesis and molecular dynamics were used to formulate a molecular mechanism for ChlG catalysis. The structure of the ChlG2HliD2 complex shows how Hlips bind to the synthase, reveals the arrangements of carotenoids and chlorophylls that mediate energy dissipation, and sheds light on the evolution of eukaryotic LHC antennae from their cyanobacterial ancestors.

plant biology↗

bioRxiv2025

The impact of carotenoid energy levels on the exciton dynamics and singlet-triplet annihilation in the bacterial Light-Harvesting 2 complex

The light-harvesting 2 (LH2) complex of purple phototrophic bacteria plays a critical role in absorbing solar energy and distributing excitation energy. Exciton dynamics within LH2 complexes are controlled by the structural arrangement and energy levels of the bacteriochlorophyll (BChl) and carotenoid (Car) pigments. However, there is still debate over the competing light-harvesting versus energy-dissipation pathways. In this work, we compared five variants of the LH2 complex from genetically modified strains of Rhodobacter sphaeroides, all containing the same BChls but different Cars with increasing conjugation: zeta-carotene (N=7; LH2Zeta), neurosporene (N=9; LH2Neu), spheroidene (N=10; LH2Spher), lycopene (N=11; LH2Lyco), and spirilloxanthin (N=13; LH2Spir). Absorption measurements confirmed that Car excited state energy decreased with increasing conjugation. Similarly, fluorescence spectra showed that the B850 BChl emission peak had an increasing red shift from LH2Zeta[->](LH2Neu/LH2Spher)[->]LH2Lyco[->]LH2Spir. In contrast, time-resolved fluorescence and ultrafast transient absorption (fs-TA) revealed similar excited state lifetimes ([~]1 ns) for all complexes except LH2Spir ([~]0.7 ns). From fs-TA analysis, an additional [~]7 ps non-radiative dissipation step from B850 BChl was observed for LH2Zeta. Further, singlet-singlet and singlet-triplet annihilation studies showed a [~]50% average fluorescence lifetime reduction in LH2Zeta at high laser power and high repetition rate, compared to [~]10-15% reductions in LH2Neu/LH2Spher/LH2Lyco and minimal lifetime change in LH2Spir. In LH2Zeta, the fastest decay component (<50 ps) became prominent at high repetition rates, consistent with strong singlet-triplet annihilation. Nanosecond TA measurements revealed long-lived (>40 s) BChl triplet states in LH2Zeta and signs of damage caused by singlet oxygen, whereas other LH2s showed faster triplet quenching ([~]18 ns) by Cars. These findings highlight a key design principle of LH2 complexes: the Car triplet energy must be significantly lower than the BChl triplet energy to efficiently quench BChl triplets that otherwise act as potent "trap states" causing exciton annihilation in laser-based experiments or photo-damage in native membranes.

biophysics↗

bioRxiv2024

Tethering ferredoxin-NADP+ reductase to photosystem I promotes photosynthetic cyclic electron transfer

Fixing CO2 via photosynthesis requires ATP and NADPH. Linear electron transfer (LET) supplies both metabolites, yet depending on environmental conditions, additional ATP is required which can be generated by cyclic electron transfer (CET). How the balance between LET and CET is set remains largely unknown. Ferredoxin(FD)-NADP+ reductase (FNR) has been suggested to act as the switch, channelling photosynthetic electrons to LET when it is bound to photosystem I (PSI) or CET when bound to cytochrome b6f. The essential role of FNR in LET precludes the use of a direct gene knock-out to test this hypothesis. We circumvented this problem by using CRISPR-Cas9 gene editing in Chlamydomonas reinhardtii to create a chimeric form of FNR tethered to PSI via PSAF. Chimeric FNR mutants exhibited impaired photosynthetic growth relative to the wild type, and decreased LET and non-photochemical quenching (NPQ) under high CO2 conditions. However, higher rates of CET and increased NPQ were found in the mutants under low CO2 or low O2 conditions where CET is normally up-regulated. Thus, rather than PSI-FNR interactions promoting LET, tethering FNR to PSI promotes CET at the expense of LET and CO2 fixation.

biochemistry↗

bioRxiv2022

Supercharged PGR5-dependent cyclic electron transfer compensates for mis-regulated chloroplast ATP synthase

The light reactions of photosynthesis couple electron and proton transfers across the thylakoid membrane, generating NADPH, and proton motive force (pmf) that powers the endergonic synthesis of ATP by ATP synthase. ATP and NADPH are required for CO2 fixation into carbohydrates by the Calvin-Benson-Bassham cycle (CBBC). The dominant {Delta}pH component of the pmf also plays a photoprotective role in regulating photosystem II (PSII) light harvesting efficiency, through non-photochemical quenching (NPQ), and cytochrome b6f (cytb6f) to photosystem I (PSI) electron transfer, via photosynthetic control. {Delta}pH can be adjusted by increasing the proton influx into the thylakoid lumen via upregulation of cyclic electron transfer (CET) or decreasing proton efflux via downregulation of ATP synthase conductivity (gH+). The interplay and relative contributions of these two elements of {Delta}pH control to photoprotection are not well understood. Here, we show that an Arabidopsis ATP synthase mutant (hope2) with 40% higher proton efflux, has supercharged CET. Double crosses of hope2 with the CET-deficient pgr5 and ndho lines reveal that PGR5-dependent CET is the major pathway contributing to higher proton influx. PGR5-dependent CET allows hope2 to maintain wild-type levels of {Delta}pH, CO2 fixation and NPQ, however photosynthetic control remains absent, and PSI is acceptor-side limited. Therefore, high CET in the absence of ATP synthase regulation is insufficient for PSI photoprotection.

plant biology↗