Light-harvesting Chlorophyll-binding Protein Research Articles

OsALB3 Is Required for Chloroplast Development by Promoting the Accumulation of Light-Harvesting Chlorophyll-Binding Proteins in Rice

ALBINO3 (ALB3) protein functions in the insertion and assembly of thylakoid membrane protein complexes and plays a critical role for chloroplast development in Arabidopsis. However, the biological function of ALB3 homologs in rice, OsALB3, remains elusive. Here, we identified a rice mutant, yellow leaf and lethal1 (yll1), that displayed yellow leaves and died at the seedling stage. The content of chlorophyll in yll1, compared with wild type, was significantly decreased. Transmission electron microscopy observation shows that the chloroplast of yll1 lacks thylakoid membranes. The causal mutation, which is located in OsALB3, was isolated by Mutmap+ combined with a simple mutation filtering process. Knockout of OsALB3 leads to yellow leaves and seedling lethality, mimicking the phenotype of yll1. OsALB3 is widely expressed and OsALB3 is chloroplast-localized. Moreover, the content of light-harvesting chlorophyll-binding proteins in yll1 is reduced. Together, our study demonstrated the essential role of OsALB3 in chloroplast development and provided clues to the possible conserved molecular function of ALB3 in rice.

Methyl jasmonate-induced senescence results in alterations in the status of chlorophyll precursors and enzymatic antioxidants in rice plants

We examined the control of chlorophyll biosynthesis and protective mechanisms during leaf senescence induced by methyl jasmonate (MeJA). After MeJA treatment, rice plants displayed evidence of great oxidative stress regarding senescence symptoms, disruption of membrane integrity, H2O2 production, and decreased chlorophyll content and photosynthetic efficiency. After 6 h of MeJA treatment, plants greatly decreased not only their levels of chlorophyll precursors, including protoporphyrin IX (Proto IX), Mg-Proto IX, Mg-Proto IX methylester, and protochlorophyllide, but also the expression levels of the chlorophyll biosynthetic genes CHLD, CHLH, CHLI, and PORB, with the greatest decreases at 78 h. MeJA-treated plants showed a noticeable degradation of light-harvesting chlorophyll-binding proteins (LHCB) at 78 h after MeJA treatment but began to downregulate expression of LHCB at 6 h. Photoprotection, as indicated by nonphotochemical quenching, slightly increased only at 6 h after MeJA treatment. In parallel to the increased activities of superoxide dismutase, catalase (CAT), ascorbate peroxidase (APX), and peroxidase, MeJA-treated plants responded to senescence by markedly upregulating the expression of APX and CAT. Our study demonstrates that rice plants developed protective mechanisms for mitigating oxidative stress by scavenging phototoxic chlorophyll precursors and activating enzymatic antioxidant responses during MeJA-induced senescence.

LHC-like Proteins: The Guardians of Photosynthesis.

The emergence of chlorophyll-containing light-harvesting complexes (LHCs) was a crucial milestone in the evolution of photosynthetic eukaryotic organisms. Light-harvesting chlorophyll-binding proteins form complexes in proximity to the reaction centres of photosystems I and II and serve as an antenna, funnelling the harvested light energy towards the reaction centres, facilitating photochemical quenching, thereby optimizing photosynthesis. It is now generally accepted that the LHC proteins evolved from LHC-like proteins, a diverse family of proteins containing up to four transmembrane helices. Interestingly, LHC-like proteins do not participate in light harvesting to elevate photosynthesis activity under low light. Instead, they protect the photosystems by dissipating excess energy and taking part in non-photochemical quenching processes. Although there is evidence that LHC-like proteins are crucial factors of photoprotection, the roles of only a few of them, mainly the stress-related psbS and lhcSR, are well described. Here, we summarize the knowledge gained regarding the evolution and function of the various LHC-like proteins, with emphasis on those strongly related to photoprotection. We further suggest LHC-like proteins as candidates for improving photosynthesis in significant food crops and discuss future directions in their research.

Client Specificity of an ATP‐independent Chaperone is Regulated by a Temperature Sensitive Switch

The chloroplast signal recognition particle 43 (cpSRP43) is an ATP‐independent chaperone that delivers the light‐harvesting chlorophyll‐binding proteins (LHCPs) to the thylakoid membrane while also maintaining the levels of tetrapyrrole biosynthesis (TBS) proteins. How cpSRP43 coordinates these roles to protect two very different classes of clients has remained unclear. We show that cpSRP43 samples two distinct conformations: a well‐folded closed state that protects unfolded membrane protein LHCPs through contacts with its substrate binding domain (SBD) and an open state with a partially disordered SBD that is less active towards LHCP but remains effective at protecting TBS proteins from heat‐induced aggregation. Binding of the additional cpSRP54 subunit stimulates cpSRP43's chaperone activity towards LHCPs by stabilizing the closed state but inhibits its chaperone activity towards TBS proteins. Strikingly, the cpSRP43‐cpSRP54 binding affinity is greatly weakened at high temperatures. Taken together, this shows that temperature can direct cpSRP43 towards one of these two client pathways. Under normal conditions, most cpSRP43 is tightly bound to cpSRP54 in the closed state and is primarily dedicated to the protection and delivery of LHCPs. However during heat stress, as chloroplast proteins become increasingly susceptible to heat‐induced aggregation, cpSRP43 dissociates from cpSRP54 and is rapidly repurposed into a stress‐responsive chaperone for TBS proteins.

SCL, Encoding a Chloroplast Signal Recognition Particle Receptor, Affects Chlorophyll Synthesis and Chloroplast Development in Rice

Crop yield is largely determined by the solar energy utilization efficiency of photosynthesis; plants with long stay-green periods have greater total photosynthetic production levels and crop yields. Here, a novel seedling chlorosis and lethality (scl) mutant exhibiting a yellow leaf and seedling-lethal phenotype was identified in rice (Oryza sativa L.). The mutant had deformed chloroplasts and almost no protein complexes in thylakoid membranes. The expression levels of photosynthesis-associated genes were significantly down-regulated in scl compared with the wild-type (WT). Positive transgenic lines generated by Agrobacterium tumefaciens-mediated transformation of the scl mutant with a complementation vector harboring SCL cDNA exhibited the normal green leaf phenotype, whereas the scl seedling harboring the empty vector displayed the yellow leaf phenotype, indicating that SCL is LOC_Os01g72800. A fusion protein expressing SCL with green fluorescent protein revealed the fluorescence signal localized to chloroplasts. The expression patterns of chloroplast development and chlorophyll biosynthesis and degradation-related genes were disordered in scl mutant, possibly resulting in the yellow leaf phenotype. These results indicated that the SCL loss of function impaired chloroplast development, chlorophyll biosynthesis, and light-harvesting chlorophyll-binding protein transportation in rice.

Chloroplast SRP43 autonomously protects chlorophyll biosynthesis proteins against heat shock.

The assembly of light-harvesting chlorophyll-binding proteins (LHCPs) is coordinated with chlorophyll biosynthesis during chloroplast development. The ATP-independent chaperone known as chloroplast signal recognition particle 43 (cpSRP43) mediates post-translational LHCP targeting to the thylakoid membrane and also participates in tetrapyrrole biosynthesis (TBS). How these distinct actions of cpSRP43 are controlled has remained unclear. Here, we demonstrate that cpSRP43 effectively protects several TBS proteins from heat-induced aggregation and enhances their stability during leaf greening and heat shock. While the substrate-binding domain of cpSRP43 is sufficient for chaperoning LHCPs, the stabilization of TBS clients requires the chromodomain 2 of the protein. Strikingly, cpSRP54-which activates cpSRP43's LHCP-targeted function-inhibits the chaperone activity of cpSRP43 towards TBS proteins. High temperature weakens the interaction of cpSRP54 with cpSRP43, thus freeing cpSRP43 to interact with and protect the integrity of TBS proteins. Our data indicate that the temperature sensitivity of the cpSRP43-cpSRP54 complex enables cpSRP43 to serve as an autonomous chaperone for the thermoprotection of TBS proteins.

A Disorder-to-Order Transition Activates an ATP-Independent Membrane Protein Chaperone

The 43 kDa subunit of the chloroplast signal recognition particle, cpSRP43, is an ATP-independent chaperone essential for the biogenesis of the light harvesting chlorophyll-binding proteins (LHCP), the most abundant membrane protein family on earth. cpSRP43 is activated by a stromal factor, cpSRP54, to more effectively capture and solubilize LHCPs. The molecular mechanism underlying this chaperone activation is unclear. Here, a combination of hydrogen–deuterium exchange, electron paramagnetic resonance, and NMR spectroscopy experiments reveal that a disorder-to-order transition of the ankyrin repeat motifs in the substrate binding domain of cpSRP43 drives its activation. An analogous coil-to-helix transition in the bridging helix, which connects the ankyrin repeat motifs to the cpSRP54 binding site in the second chromodomain, mediates long-range allosteric communication of cpSRP43 with its activating binding partner. Our results provide a molecular model to explain how the conformational dynamics of cpSRP43 enables regulation of its chaperone activity and suggest a general mechanism by which ATP-independent chaperones with cooperatively folding domains can be regulated.

Protein Sorting within Chloroplasts.

Chloroplasts have multiple suborganellar membranes. Correct and efficient translocation of chloroplast proteins from their site of synthesis into or across membranes to their functional compartments are fundamental processes. In recent years, several new components and regulatory mechanisms involved in chloroplast protein import and sorting have been explored. Moreover, the formation of liquid-liquid phase transition (LLPT) has been recently reported as a novel mechanism for regulating chloroplast protein sorting. Here, we overview the recent advances of both nuclear- and chloroplast-encoded protein trafficking to their final destination within chloroplasts, and discuss the novel components and regulatory mechanisms of intrachloroplast sorting. Furthermore, we propose that LLPT may be a universal and conserved mechanism for driving organelle protein trafficking and organelle biogenesis.

Modulation of chloroplast components and defense responses during programmed cell death in tobacco infected with Pseudomonas syringae

We examined how tobacco plants coordinate chloroplast components and defense responses during Pseudomonas syringae pv. tomato (Pst) infection. Tobacco leaves infiltrated with Pst induced weak necrosis at 24 h post-infiltration (hpi) and severe necrosis at 48 hpi. Membrane damage, as shown by cellular leakage and malondialdehyde, and H2O2 production began to increase at 12 hpi and continuously increased at 24–72 hpi in Pst-infiltrated leaves. Pst infection resulted in decreases in light-harvesting chlorophyll-binding proteins (Lhc), Lhcb transcripts, electron transport rate, and Fv/Fm, indicating the impairment in structure and function of photosystem II. Photochemical quenching, qP, continuously decreased in Pst-infiltrated leaves at 24–48 hpi, whereas nonphotochemical quenching, NPQ, exhibited a 2-fold increase at 24 hpi and a decrease at 48 dpi. In response to Pst infection, chlorophyll began to decrease at 48 hpi, whereas levels of protoporphyrin IX (Proto IX), Mg-Proto IX, Mg-Proto methylester, and protochlorophyllide drastically decreased or disappeared as early as 24 hpi. Pst-infiltrated leaves greatly up-regulated the expression of ROS scavenging genes, Fe-SOD, APX, and CAT1, as well as defense-related genes, PII, PR1, PR2, PALa, and CHS1. Our study suggests that the modulation of photosynthetic components during pathogen infection, particularly in relation to the fast degradation of photosensitizing porphyrin intermediates and the increase in photoprotective NPQ, may contribute to attenuating cellular damage in the early stages of programmed cell death induced by Pst.

English

A glasshouse experiment was conducted to compare the performance of a stay-green QTL introgression line (RSG 03123) and its parental lines (B35, a stay-green and R16, a high yielding cultivar with rapid senescence) after flowering, when water-limited (WL) or well-watered (WW). Flowering time and plant height were similar in RSG 03123 and the recurrent parent, R16. Under WW conditions, retention of green leaf area (GLA) and chlorophyll was similar in RSG 03123 and R16, whereas under WL conditions RSG 03123 retained significantly (P<0.05) more GLA. The rate of senescence was also lower in RSG 03123 as compared to R16 under WL conditions. B35 displayed a delayed onset of senescence under both WW and WL conditions while both R16 and RSG 01323 displayed an earlier onset of senescence under WL conditions. Photochemical efficiency of photosystem II (ΦPSII), gas exchange characteristics, and total soluble carbohydrates contents, in both leaf and stem, were higher in RSG 03123 than in R16 under WL conditions. These parameters were enhanced in RSG 03123 due to higher retention of chloroplast proteins, such as light-harvesting chlorophyll binding protein complex of PSII (LHCPII), oxygen-evolving complex 33 kDa (OEC33), phosphoenolpyruvate carboxylase (PEPC) and the large subunit of ribulose bisphosphate carboxylase-oxygenase (Rubisco). The stay-green characteristics of RSG 03123 were intermediate between R16 and B35; nonetheless, the results show that stay-green QTL are functional during senescence and improve tolerance to water limitation after flowering. Key words: Stay-green QTL, introgression, drought, photosynthesis.

LIL3, a Light-Harvesting Complex Protein, Links Terpenoid and Tetrapyrrole Biosynthesis in Arabidopsis thaliana.

The LIL3 protein of Arabidopsis (Arabidopsis thaliana) belongs to the light-harvesting complex (LHC) protein family, which also includes the light-harvesting chlorophyll-binding proteins of photosystems I and II, the early-light-inducible proteins, PsbS involved in nonphotochemical quenching, and the one-helix proteins and their cyanobacterial homologs designated high-light-inducible proteins. Each member of this family is characterized by one or two LHC transmembrane domains (referred to as the LHC motif) to which potential functions such as chlorophyll binding, protein interaction, and integration of interacting partners into the plastid membranes have been attributed. Initially, LIL3 was shown to interact with geranylgeranyl reductase (CHLP), an enzyme of terpene biosynthesis that supplies the hydrocarbon chain for chlorophyll and tocopherol. Here, we show another function of LIL3 for the stability of protochlorophyllide oxidoreductase (POR). Multiple protein-protein interaction analyses suggest the direct physical interaction of LIL3 with POR but not with chlorophyll synthase. Consistently, LIL3-deficient plants exhibit substantial loss of POR as well as CHLP, which is not due to defective transcription of the POR and CHLP genes but to the posttranslational modification of their protein products. Interestingly, in vitro biochemical analyses provide novel evidence that LIL3 shows high binding affinity to protochlorophyllide, the substrate of POR. Taken together, this study suggests a critical role for LIL3 in the organization of later steps in chlorophyll biosynthesis. We suggest that LIL3 associates with POR and CHLP and thus contributes to the supply of the two metabolites, chlorophyllide and phytyl pyrophosphate, required for the final step in chlorophyll a synthesis.

SNOWY COTYLEDON 2 Promotes Chloroplast Development and Has a Role in Leaf Variegation in Both Lotus japonicus and Arabidopsis thaliana

Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 (SCO2) is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCB1. In Arabidopsis thaliana, SCO2 function was previously reported to be restricted to cotyledons. Here we show that disruption of SCO2 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale-green true leaves can also be observed in A. thaliana sco2 (atsco2) mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSII–LHCII complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant development and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSII assembly or repair and constitutes a novel factor involved in leaf variegation.

Domain Organization in the 54-kDa Subunit of the Chloroplast Signal Recognition Particle

Chloroplast signal recognition particle (cpSRP) is a heterodimer composed of an evolutionarily conserved 54-kDa GTPase (cpSRP54) and a unique 43-kDa subunit (cpSRP43) responsible for delivering light-harvesting chlorophyll binding protein to the thylakoid membrane. While a nearly complete three-dimensional structure of cpSRP43 has been determined, no high-resolution structure is yet available for cpSRP54. In this study, we developed and examined an in silico three-dimensional model of the structure of cpSRP54 by homology modeling using cytosolic homologs. Model selection was guided by single-molecule Förster resonance energy transfer experiments, which revealed the presence of at least two distinct conformations. Small angle x-ray scattering showed that the linking region among the GTPase (G-domain) and methionine-rich (M-domain) domains, an M-domain loop, and the cpSRP43 binding C-terminal extension of cpSRP54 are predominantly disordered. Interestingly, the linker and loop segments were observed to play an important role in organizing the domain arrangement of cpSRP54. Further, deletion of the finger loop abolished loading of the cpSRP cargo, light-harvesting chlorophyll binding protein. These data highlight important structural dynamics relevant to cpSRP54’s role in the post- and cotranslational signaling processes.

Regulation of Structural Dynamics within a Signal Recognition Particle Promotes Binding of Protein Targeting Substrates

Protein targeting is critical in all living organisms and involves a signal recognition particle (SRP), an SRP receptor, and a translocase. In co-translational targeting, interactions among these proteins are mediated by the ribosome. In chloroplasts, the light-harvesting chlorophyll-binding protein (LHCP) in the thylakoid membrane is targeted post-translationally without a ribosome. A multidomain chloroplast-specific subunit of the SRP, cpSRP43, is proposed to take on the role of coordinating the sequence of targeting events. Here, we demonstrate that cpSRP43 exhibits significant interdomain dynamics that are reduced upon binding its SRP binding partner, cpSRP54. We showed that the affinity of cpSRP43 for the binding motif of LHCP (L18) increases when cpSRP43 is complexed to the binding motif of cpSRP54 (cpSRP54pep). These results support the conclusion that substrate binding to the chloroplast SRP is modulated by protein structural dynamics in which a major role of cpSRP54 is to improve substrate binding efficiency to the cpSRP.

GTP Hydrolysis is Not Required for Post‐translational Protein Insertion by a Chloroplast Signal Recognition Particle

Light harvesting chlorophyll binding proteins (LHCPs) are a family of nuclear encoded chloroplast thylakoid proteins that are synthesized in the cytosol, imported into the chloroplast stroma, and subsequently directed to the thylakoid membrane. LHCP targeting/insertion into thylakoid membranes requires GTP, a thylakoid protein insertase, and two prokaryote derived GTPases, chloroplast signal recognition particle (cpSRP) and its membrane receptor (cpFtsY). cpSRP binds LHCP in the stroma to form a soluble targeting complex that is directed to thylakoids owing to affinity of cpSRP for cpFtsY, an interaction requiring GTP. LHCP insertion requires Albino‐3 (Alb3), which stimulates GTP hydrolysis by cpSRP/cpFtsY and releases cpSRP from cpFtsY. The role of GTP in LHCP insertion is less clear. Using assays that reconstitute LHCP integration into isolated thylakoids, we have demonstrated that GTP hydrolysis is not required for LHCP insertion. GMP‐PNP, a non‐hydrolysable analogue of GTP, is sufficient to support LHCP integration. Surprisingly, LHCP integration supported by GMP‐PNP, and not GTP, is sensitive to elevated concentrations of free cpSRP/cpFtsY. Our data supports a mechanism in which guanine nucleotide binding by cpSRP/cpFtsY is required for LHCP release from cpSRP whereas GTP hydrolysis serves to recycle cpSRP from cpFtsY for subsequent rounds of targeting. It is hypothesized that guanine nucleotide binding will only serve to release LHCP from cpSRP in the presence of an available Alb3 insertase to prevent unproductive targeting.

Three-Dimensional Structure of the 54-Kda Subunit of the Chloroplast Signal Recognition Particle using Molecular Modeling

The chloroplast signal recognition particle is a heterodimeric complex of the 54kDa cytosolic signal recognition particle homologue (cpSRP54), and a novel 43kDa subunit (cpSRP43). While a nearly complete three-dimensional structure of cpSRP43 has been obtained, no structure is yet available for cpSRP54. The three-dimensional structure for cpSRP54 could provide valuable information for the rationalization of the extensive information already available regarding its function, and in the understanding of the as yet undetermined mechanism of light harvesting chlorophyll binding protein's (LHCP) insertion into the thylakoid membrane. In this study, we developed an in silico, three-dimensional structural model of cpsRP54 using a combination of homology modeling, de novo structure prediction and molecular dynamics simulation. The resulting structure is consistent with the known properties of the protein and sheds new light on some of the mechanistic details of its functioning.

C-Terminal M4 Peptide Fragment of Albino 3 Interaction with cpSRP-43

The protein complexes responsible for insertion of light harvesting chlorophyll binding protein into the thylakoid membrane of the chloroplast are the concern of this investigation. Critical for this insertion, the membrane bound protein Albino 3 has been shown to bind a cpSRP-FTSY-LHCP complex. The following study implicates the stromal, c-terminus of Albino 3 as an imperative binding interaction in this process. Isothermal titration calorimetry and 1H-15N HSQC were used to understand the binding of a 16 residue segment from this region of Albino 3 known as M4. The data show that this segment binds with 14uM affinity and that electrostatic interactions with E327 and E356 on the 43kDa subunit of cpSRP are essential for this event. Importantly, significant binding is not observed when two domains of cpSRP-43, CD2 and CD3, are separated. Far UV and intrinsic fluorescence experiments also show that, upon M4 binding, moderate conformational changes in CD2-CD3 occur.

Investigation of the Structural Stability of cpSRP43 Chromodomain2 by Hydrogen-Deuterium Exchange

The essential pathway by which light-harvesting chlorophyll binding proteins are inserted into the thylakoid of the chloroplast is mediated by a unique signal recognition particle (cpSRP). Its novel 43kDa subunit (cpSRP43) contains three chromodomains (CDs). CD2 has been shown to be significant in various aspects of the cpSRP-mediated pathway. While structural information about CD2 is known, this study aims to gain a more comprehensive understanding of the structural stability through analysis of hydrogen-deuterium exchange. 15N-labeled CD2 samples will be produced from over-expression of CD2 in E. coli cultured in labeled media. Subsequently, the readily-occurring exchange between amide hydrogen and solvent deuterium will be monitored by NMR spectroscopy. The kinetics of the exchange can provide useful information about the free energy of exchange and thus the stability.

Binding of chloroplast signal recognition particle to a thylakoid membrane protein substrate in aqueous solution and delineation of the cpSRP43–substrate interaction domain

A cpSRP [chloroplast SRP (signal recognition particle)] comprising cpSRP54 and cpSRP43 subunits mediates the insertion of light-harvesting proteins into the thylakoid membrane. We dissected its interaction with a full-length membrane protein substrate in aqueous solution by insertion of site-specific photo-activatable cross-linkers into in vitro-synthesized Lhcb1 (major light-harvesting chlorophyll-binding protein of photosystem II). We show that Lhcb1 residues 166-176 cross-link specifically to the cpSRP43 subunit. Some cross-link positions within Lhcb1 are in the 'L18' peptide required for targeting of cpSRP substrates, whereas other cross-linking positions define a new targeting signal in the third transmembrane span. Lhcb1 was not found to cross-link to cpSRP54 at any position, and cross-linking to cpSRP43 is unaffected by the absence of cpSRP54. cpSRP43 thus effectively binds substrates autonomously, and its ability to independently bind an extended 20+-residue substrate region highlights a major difference with other SRP typeswhere the SRP54 subunit binds to hydrophobic target sequences. The results also show that cpSRP43 can bind to a hydrophobic, three-membrane span, substrate in aqueous solution, presumably reflecting a role for cpSRP in the chloroplast stroma. This mode of action, and the specificity of the cpSRP43-substrate interaction, may be associated with cpSRP's unique post-translational mode of action.

LTD is a protein required for sorting light-harvesting chlorophyll-binding proteins to the chloroplast SRP pathway

Higher plants require chloroplasts for essential functions in photosynthesis and other important physiological processes, such as sugar, lipid and amino-acid biosynthesis. Most chloroplast proteins are nuclear-encoded proteins that are synthesized in the cytosol as precursors, and imported into chloroplasts by protein translocases in the outer and inner chloroplast envelope. The imported chloroplast proteins are then translocated into or across the thylakoid membrane by four distinct pathways. However, the mechanisms by which the imported nuclear-encoded proteins are delivered to these pathways remain largely unknown. Here we show that an Arabidopsis ankyrin protein, LTD (mutation of which causes the light-harvesting chlorophyll-binding protein translocation defect), is localized in the chloroplast and using yeast two-hybrid screens demonstrate that LTD interacts with both proteins from the signal recognition particle (SRP) pathway and the inner chloroplast envelope. Our study shows that LTD is essential for the import of light-harvesting chlorophyll-binding proteins and subsequent routing of these proteins to the chloroplast SRP-dependent pathway.