Plant Cells: Peroxisomes and Glyoxysomes

Robert Paul Donaldson, The George Washington University, Washington DC, USA Masoumeh Assadi, The George Washington University, Washington DC, USA Konstantina Karyotou, The George Washington University, Washington DC, USA Tulin Olcum, The George Washington University, Washington DC, USA Tianqing Qiu, The George Washington University, Washington DC, USA
Peroxisomes and glyoxysomes are membrane enclosures, both referred to as microbodies, which contain oxidative enzymes that participate in photorespiration in leaves, nitrogen metabolism in root nodules, and fat conversions in seeds. The enzymes found within microbodies are brought in from the cytosol by information described as peroxisomal targeting sequences (PTS).

Secondary article
Article Contents
. Basic Structure, Basic Functions . Photorespiration: Leaf Peroxisomes . Fixed Nitrogen Conversion into Ureides: Root Nodule Peroxisomes . Breakdown of Fatty Acids during Germination: Glyoxysomes . Peroxisome Formation: Glyoxysome Peroxisome Conversions . Conclusion

Basic Structure, Basic Functions

A peroxisome or a glyoxysome consists of a specic group of enzymes or proteins enclosed by a single membrane. These organelles, which are in the range of 1 mm in diameter, are visible under the electron microscope and are sometimes referred to as microbodies. In higher plants, at least four classes of peroxisomes have been identied: glyoxysomes, leaf peroxisomes, root nodule peroxisomes and unspecialized peroxisomes. All classes of peroxisomes have the following characteristics: (1) they have a single membrane; (2) they have high equilibrium density of c. 1.25 g cm 2 2 in sucrose gradient centrifugation; and (3) their matrix (internal content) is nely granular. Although all classes possess the common characteristics, they have distinct metabolic roles specied by the developmental stage and type of cell. Catalase, which is by denition always found in these organelles, can be stained black such that the organelles are more obvious in electron micrographs. The proteins within this type of organelle are visible as a granular matrix somewhat more dense than that of the cytosol. This type of organelle does not have any internal membranous structures but in some cases the matrix includes a striking proteinaceous crystal or dense aggregate, visible in electron microscopy. Peroxisomes or glyoxysomes can also be visualized in uorescence microscopy using antibodies specic to one of their proteins, such as catalase. The relatively simple structure of the internal matrix of microbodies distinguishes them from chloroplasts or mitochondria, which have internal membranes that are folded or stacked. In the photosynthetic cells of leaves the peroxisomes are often in contact with chloroplasts and mitochondria; these three organelles interact with each other in photorespiration. Glyoxysomes are found in contact with lipid bodies in cotyledons or

endosperm where fatty acids are being converted to carbohydrate (sugars) during germination. Images of whole plant cells indicate that there may be a few hundred microbodies in a cell. In some instances the microbodies may be tubular or interconnected and appear to be dividing. All four known classes of microbodies found in plant cells are organelles that, by denition, contain activities that produce and destroy hydrogen peroxide (H2O2), which is a toxic agent. Glyoxysomes are specialized peroxisomes that are present in postgerminative seedlings of oil seeds and senescent organs. Glyoxysomes are involved in storage lipid mobilization in growing seedlings via the glyoxylate cycle. Succinate produced in glyoxysomes is ultimately converted to sucrose in the cytosol. It is presumed that presence of glyoxysomes in senescent organs is in response to the mobilization of membrane lipids. Leaf peroxisomes are present in green and photosynthetically active tissues, such as green cotyledons and leaves. These peroxisomes contain enzymes that are required for the light-dependent process of photorespiration. Root nodule peroxisomes are present in the root nodules of certain legumes and involved in nitrogen metabolism. In many tropical legumes, nitrogen is transported in the form of ureides, allantoin and allantoic acid. Reactions of ureide biosynthesis take place in several subcellular compartments. One of the last steps of this pathway, the conversion of urate to allantoin, is catalysed by urate oxidase in peroxisomes. Unspecialized peroxisomes are present in plant tissues that are not photosynthetically active and that lack storage lipids, such as the roots of most plants. Unspecialized peroxisomes can be distinguished from other forms of peroxisomes by their small size, low frequency and density
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Plant Cells: Peroxisomes and Glyoxysomes

compared to glyoxysomes and leaf-type peroxisomes. Their specic role in the cellular metabolism is not known. The metabolic processes that take place in peroxisomes often bypass energy conservation steps. The prototypical enzyme of these organelles is an oxidase that generates H2O2, such as the glycolate oxidase (GO) of leaf peroxisomes or the fatty acylCoA oxidase (AO) of glyoxysomes. These oxidases contain avin (as FAD), which accepts two hydrogens from a substrate (e.g. glycolate or acylCoA) and transfers them to oxygen, resulting in H2O2. In the mitochondria this process would be coupled to energy conservation, where the hydrogens recovered from the substrate serve as a source of electrons to power mitochondrial ATP generation. The advantage of directly transferring the hydrogens to oxygen in peroxisomes is that the metabolic processes can take place even when the cell is not consuming ATP and when additional ATP does not need to be generated.

Photorespiration: Leaf Peroxisomes

Photorespiration and photosynthesis are opposing processes that occur in the cells of leaves or other green tissues. Both processes are initiated in chloroplasts, but photorespiration involves a diversion into leaf peroxisomes and mitochondria. As a cell of a young leaf expands, its vacuole lls with water and spreads the cytoplasm around the periphery of the cell. In the cytoplasm the chloroplasts, peroxisomes and mitochondria become loaded with the enzymes needed to absorb light and carbon dioxide to create new organic molecules for the rest of the plant. The carbon dioxide is taken in by the chloroplast enzyme, ribulose-1,5-bisphosphate carboxylase (Rubisco), the most abundant enzyme in the cell, and is normally assimilated into the three-carbon molecule phosphoglyceric acid (PGA), which is used to synthesize sugars and other organic molecules. Photorespiration commences when oxygen replaces carbon dioxide in Rubisco. This results in the two-carbon molecule, phosphoglycolate, instead of PGA. The phosphoglycolate can be recycled back into PGA by a circuitous process through leaf peroxisomes, mitochondria and chloroplasts with the use of oxygen and the loss of one carbon in four as carbon dioxide, hence the designation photorespiration. The glycolate, relieved of its phosphate, is passed from a chloroplast to a peroxisome. Here it is subject to a typical peroxisomal enzyme, glycolate oxidase, which transfers two hydrogens to oxygen, resulting in hydrogen peroxide, which is broken down by catalase. The result is glyoxylate, which accepts an amino group (NH2) to become the amino acid glycine. This is transported into the mitochondria. In a photosynthetic cell the proteins of photorespiration are the most abundant in the mitochondria. Here a complex of four proteins
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combines two molecules of glycine to create a molecule of serine with the release of a carbon dioxide molecule and an amino group. The serine returns to a peroxisome where additional enzymes complete its conversion to glycerate by transferring its amino group to glyoxylate, followed by reduction by hydroxypyruvate reductase using NADH2. The glycerate re-enters a chloroplast where it is phosphorylated, nally yielding PGA. These processes require the transport of metabolites through the membranes of the various organelles including the peroxisomes. There may be selective channel proteins in the membranes that regulate the transport of metabolites. A porin protein discovered in the membranes of peroxisomes may represent such a channel (Reumann et al., 1998). The process of photorespiration is very signicant in plants and becomes especially important when leaf stomata close to reduce water loss. Then the supply of carbon dioxide within the leaf diminishes as it is assimilated and at the same time the concentration of oxygen increases, favouring photorespiration. The carbon dioxide released by photorespiration can be reassimilated by Rubisco, thus allowing use of the light energy absorbed by the chloroplast. Although photorespiration is counterproductive to photosynthesis, it may be necessary to protect the leaf cells from damage due to light absorption.

Fixed Nitrogen Conversion into Ureides: Root Nodule Peroxisomes

Root nodule peroxisomes of certain tropical legumes synthesize allantoin, which serves as the major metabolite for nitrogen transport in these plants. The ureides, allantoin and allantoic acid, are the predominant form of nitrogen transported in the xylem of soya bean and cowpea plants growing symbiotically. The synthesis of allantoic acid presumably derives from the degradation of purines. Urate oxidase (UO), one enzyme in the purine degradation pathway, is normally found in peroxisomes, along with catalase, which consumes the hydrogen peroxide produced by UO. Uric acid is oxidized by UO to allantoin within peroxisomes. Small amounts of UO are present in glyoxysomes of germinating oil seeds and of potato tubers, while traces of UO are also present in peroxisomes from other plant tissues. In all cases UO is easily solubilized and is not part of the crystalline core of the peroxisome. Allantoinase and allantoicase, enzymes participating in the biogenesis of allantoin and allantoic acid, have been reported to be present in peroxisomes from amphibian and sh livers. Approximately one-half of the allantoinase activity in castor bean endosperm is associated with glyoxysomes; the remainder is in the proplastids (Hanks et al., 1981).

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Plant Cells: Peroxisomes and Glyoxysomes

Breakdown of Fatty Acids during Germination: Glyoxysomes

Metabolically, plant and animal cells dier in many important respects. In particular, plant cells, along with some microorganisms, can carry out the net synthesis of carbohydrate from fat. This conversion is crucial in the development of seeds, in which a great amount of energy is stored in the form of triacylglycerols. As such seeds germinate, triacylglycerol stored in lipid bodies is broken down, transported to glyoxysomes and eventually converted to sugars, which provide energy and the raw material needed for growth of the plant. In contrast, animal cells cannot carry out the net synthesis of carbohydrate from fat. In plants, catabolism of triacylglycerols in the lipid bodies yields fatty acids and glycerol. Fatty acids undergo b-oxidation to yield acetylCoA, which can then be incorporated into carbohydrate through the glyoxylate cycle. The conversion of triacylglycerols into sugars involves metabolism in glyoxysomes. While leaf peroxisomes have a key role in photorespiration, glyoxysomes are the sites of b-oxidation of fatty acids and the glyoxylate cycle (Cooper and Beevers, 1969). These pathways are essential to the maintenance of gluconeogenesis initiated by the degradation of reserve or structural lipids. bOxidation of fatty acids occurs in most plant microbodies, but has a more important function in those organelles from fat-storing tissues of oilseeds. In these organelles, fatty acids are degraded via the b-oxidation pathway to acetyl CoA, which in turn is metabolized by the glyoxylate cycle to succinate, bypassing the decarboxylating steps of the Krebs cycle (Tolbert, 1981). Succinate is then used for gluconeogenesis or synthesis of other metabolic intermediates. The glyoxysomal b-oxidation of fatty acids is a recurring sequence of four reactions shown in Figure 1: oxidation (dehydrogenation) by acylCoA oxidase (AO), hydration by enoylCoA hydratase combined with a second oxidation by 3-hydroxy acylCoA dehydrogenase, both catalysed within a bifunctional protein (BP), and nally thiolysis by 3-ketoacylCoA thiolase (TH). The FADlinked acylCoA oxidase transfers electrons not to the respiratory electron transport chain but directly to oxygen, without recovery of chemical energy (ATP). The oxygen is reduced to hydrogen peroxide, which in turn is scavenged by catalase (CAT). The dehydrogenase produces NADH2 and the thiolase uses CoA to remove the last two carbons of the 3-ketoacylCoA to yield acetylCoA. As stored lipids are metabolized during seed germination, the acetylCoA produced by b-oxidation in glyoxysomes is transferred to the glyoxylate cycle, which can be considered as an anabolic variant of the citric acid cycle. The glyoxylate cycle converts two molecules of acetyl CoA into one molecule of succinate, as shown in Figure 1.

This involves two glyoxylate cycle-specic enzymes, namely isocitrate lyase (IL) and malate synthase (MS), and three enzyme activities similar to those from the citric acid cycle, namely citrate synthase (CS), aconitase (AC) and malate dehydrogenase (MD). Since the glyoxylate pathway bypasses the two reactions of the Krebs cycle where carbon is lost, each turn of the cycle involves incorporation of two two-carbon molecules and results in the net synthesis of the four-carbon molecule, succinate. This is transported from the glyoxysome to the mitochondria where it is converted through the Krebs cycle to oxalacetate, which is readily utilized by gluconeogenesis for carbohydrate synthesis. The reduced cofactors that are produced during both boxidation and glyoxylate cycle, namely NADH2 and FADH2, do not have direct access to the mitochondrial electron transport system. They must therefore be reoxidized in order for both pathways to remain functional. The acylCoA oxidase of glyoxysomal b-oxidation avoids that by transferring electrons from the FADH2 directly to oxygen, resulting in hydrogen peroxide. Hydrogen peroxide is produced in abundance within glyoxysomes during this process or from the disproportionation of superoxide radicals by superoxide dismutase. Superoxide radicals can be produced by the transfer of electrons from NADH2 to oxygen via a protein in the membrane (Del Rio et al., 1992). The hydrogen peroxide is decomposed either by catalase (CAT) inside the glyoxysome or by an ascorbate-specic peroxidase (AP) present at the glyoxysomal membrane. NADH2 produced by the 3-hydroxy acylCoA dehydrogenase and by malate dehydrogenase in the glyoxylate cycle also accumulates within glyoxysomes, and is oxidized by the electron transport proteins in the glyoxysomal membrane. These proteins include ascorbate peroxidase (AP), ascorbate free radical reductase (AR) and, possibly, cytochrome b5 and glutathione reductase. Ascorbate peroxidase utilizes hydrogen peroxide to catalyse a one-electron oxidation of ascorbate, resulting in the formation of ascorbate free radicals. Regeneration of ascorbate is achieved by ascorbate free radical reductase (AR), using NADH2 as an electron donor. Overall, glyoxysomal metabolism results in the production of a 2 ., H2O2 and variety of reactive species, such as O2 ascorbate free radicals. At the same time the glyoxysomes include the appropriate detoxifying enzymes, such as catalase and the enzymes located in the membrane AP and AR, which can protect against cell damage (Bunkelmann and Trelease, 1996; Ishikawa et al., 1998).

Peroxisome Formation: Glyoxysome Peroxisome Conversions

In the oil-storing cotyledons of seeds such as cotton, cucumber or legumes, a population of glyoxysomes is
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Plant Cells: Peroxisomes and Glyoxysomes

Lipid body Asc Triacyl glycerols Glycerol Fatty acids H 2O O2 Fatty acid Glyoxysome
AcylCoA CAT

Asc AP AR
As c

H 2O 2 FAD AO

H 2O BP
3-OH-acylCoA

EnoylCoA

Asc AR Asc

-Oxidation

BP

NAD+

NADH2 3-ketoacylCoA CoA-SH TH AcetylCoA

Succinate Isocitrate AC Glyoxylate cycle Succinate Citrate CS Oxaloacetate AcetylCoA Mitochondrion

IL

Glyoxylate MS Malate MD

NAD+ NADH2 AR Cytosol

Figure 1 b-Oxidation and glyoxylate cycle enzymes in glyoxysomes. The b-oxidation enzymes are acyl CoA oxidase (AO), enoyl CoA hydratase combined with 3-hydroxy acyl CoA dehydrogenase in the bifunctional protein (BP) and 3-ketoacyl CoA thiolase (TH). Glyoxylate-cycle enzymes are isocitrate lyase (IL) and malate synthase (MS), citrate synthase (CS), aconitase (AC) and malate dehydrogenase (MD). Membrane enzymes include ascorbate peroxidase (AP) and ascorbate free radical reductase (AR). Both catalase (CAT) and AP consume hydrogen peroxide. Asc, ascorbate; Asc.; ascorbate free radical.

converted into a population of leaf peroxisomes following exposure to light, resulting in greening of the tissue. The functions of the microbodies are thus converted from lipid metabolism to photorespiratory metabolism. Two ideas, the one-population and the two-population hypotheses, have been proposed for the interconversions and origins of specialized peroxisomes. According to the rst hypothesis, leaf peroxisomes are formed from existing glyoxysomes by insertion of newly synthesized leaf peroxisome-specic enzymes and depletion of glyoxysomal-specic enzymes. In contrast, the second hypothesis suggests de novo formation of glyoxysomes and leaf peroxisomes. According to the one-population hypothesis, glyoxysomal-specic enzymes and leaf peroxisome-specic enzymes are present in single microbody species throughout seedling growth, even after illumination. The second hypothesis suggests that glyoxysomes and peroxisomes contain completely dierent enzymes. However, several lines of evidence support the rst hypothesis. For example, microbodies with both glyoxysomal-specic and leaf peroxisomalspecic enzymes have been identied during the transitional stage, using immunocytochemical analysis. This indicates that glyoxysomes are directly transformed to leaf
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peroxisomes during greening. Additional support for this idea comes from the same kind of nding during senescence of cotyledons or leaves, a stage in which leaf peroxisomes are converted to glyoxysomes and the stores of carbon and nitrogen are transferred to newly developing tissues. Immunocytochemical analysis revealed that enzymes specic to glyoxysomes and to leaf peroxisomes are both present in microbodies of senescing cotyledons (Titus and Becker, 1985). Although the morphological appearance of microbodies in cotyledons is the same during transitions from glyoxysomes to peroxisomes, their enzymatic contents are changed drastically. As discussed above, each specialized microbody contains dierent enzymes. Activities of glyoxysomal-specic enzymes, such as malate synthase and citrate synthase, increase with germination and decrease gradually after lipid stores are depleted. At this stage, activities of leaf peroxisome-specic enzymes are at the lowest level. Rapid increase in activities of leaf peroxisome-specic enzymes and decrease in activities of glyoxysomal-specic enzymes occurs when seedlings are transferred to the light. The transition of glyoxysomes to leaf peroxisomes and the accumulations of new proteins in

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Plant Cells: Peroxisomes and Glyoxysomes

microbodies can be regulated in several ways: through gene expression, protein transport, mRNA splicing and protein degradation. These are discussed below.

Regulation of gene expression

The mRNA levels for glyoxysomal-specic enzymes malate synthase and citrate synthase increase rapidly during germination in the dark and decline markedly after exposure of the cotyledons to the light. Expression of the malate synthase gene is inhibited by sucrose, the end product of the metabolism of stored lipid. On the other hand mRNA levels of leaf peroxisome enzymes such as glycolate oxidase are low during germination in the dark and increase rapidly during greening. Regulation of leaf peroxisome enzymes is light dependent and it has been reported that phytochrome plays a role in signal transduction. Accordingly, it is suggested that light and levels of metabolites are regulatory factors for microbody transition events.

Regulation of protein transport into microbodies

All microbody enzymes are synthesized in the cytoplasm and transported into microbodies posttranslationally. The enzymes include certain sequences of amino acids that serve as targeting information. Only two major types of microbody targeting signals are known in plants as well as in mammals, insects, fungi and protists. The peroxisomal targeting signal-1, PTS-1, is a C-terminal tripeptide such as Ser-Lys-Leu (SKL) that is present in mature proteins in microbodies. Conservative variations in the amino acids of the PTS-1 sequence can be tolerated without loss of targeting activity. For example, the targeting sequences are SRL for malate synthase from castor bean, PRL for glycolate oxidase, and ARM or SRM for isocitrate lyases. However, the removal of the ARM from castor bean isocitrate lyase does not stop import of the protein, suggesting that there is additional targeting information in the protein (Gao et al., 1996). Experimental alterations of the PTS-1 suggest that other amino acids may be functional in the tripeptide and that additional amino acids nearby may also contribute to recognition (Mullen et al., 1997a; Wolins and Donaldson, 1997). The C-terminal sequence of cotton seed glyoxysomal catalase is -NVKPSI and the experimental evidence indicates that some of the amino acids in addition to the PSI are necessary for import (Mullen et al., 1997b). Many other proteins from a variety of plant species (see Table 1) t the rather relaxed PTS-1 consensus, but in the absence of experimental evidence it cannot be assumed that each of these sequences functions as a PTS-1. Comparisons of the amino acid sequences from several enzymes indicate that each enzyme has a particular version of the PTS-1 that is found in several species. A

cytosolic PTS-1 receptor that interacts with a peroxisomal membrane docking protein has been described for human and yeast peroxisomes. Some in vitro studies indicate that a similar receptor exists in plants (Wolins and Donaldson, 1994; Brickner et al., 1997; Kragler et al., 1998). Most of the microbody proteins have the C-terminal PTS-1, but a few have a second type of targeting signal near their N-terminal, PTS-2. This consists of a sequence such as RLXXXXXHL, where X can be any amino acid (Gietl et al., 1994). These proteins include glyoxysomal 3ketoacylCoA thiolase, malate dehydrogenase and citrate synthase, which are synthesized with larger molecular mass in the cytosol. Their N-terminal PTS-2 peptides are then cleaved upon the targeting of the enzymes into microbodies. Experiments showed that a fusion protein composed of the N-terminal region of glyoxysomal citrate synthase was transported to glyoxysomes, leaf peroxisomes and unspecialized microbodies and was subsequently processed. This suggests that microbodies use the same transport mechanism and that dierentiation of microbodies is not regulated at the level of recognition of the targeting information. It has been observed that proteins that have had their targeting information removed experimentally are imported into glyoxysomes if accompanied by proteins that do have the targeting information (Lee et al., 1997). The implication is that the protein lacking a PTS can piggyback or associate with the protein having the PTS, and that the two proteins can enter together. How such an assemblage would traverse the membrane of the glyoxysome is not understood.

Regulation of mRNA splicing

Hydroxypyruvate reductase (HPR) is one of the leaf peroxisome-specic enzymes that is induced and accumulates in microbodies during greening. cDNA analysis of pumpkin cotyledons showed that two very similar cDNAs encode for this enzyme. The only dierence between the two encoded proteins is that HPR-1 contains PTS-1 and HPR-2 does not. Genomic DNA analysis suggested that the HPR-1 and HPR-2 are encoded from the same gene by alternative splicing. Accumulation during greening of HPR-1 and HPR-2, in leaf peroxisomes and the cytosol, respectively, suggests that alternative mRNA splicing may play a regulatory role in microbody transition (Hayashi et al., 1996).

Regulation at the level of protein degradation

During transition of glyoxysomes to leaf peroxisomes, de novo synthesis of glyoxysomal-specic enzymes is prevented by depletion and degradation of mRNA for these enzymes. Furthermore, preexisting glyoxysomal-specic enzymes are degraded by proteolytic enzymes present in
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Plant Cells: Peroxisomes and Glyoxysomes

Table 1 Peroxisomal targeting sequences (PTS-1 and PTS-2) Protein Malate synthase Species Cucumbera Soya beana Rapea Castor beana Spinach Arabidopsisa Cucumber Rice Tomato Soya beana Castor beana Cucurbita Arabidopsis Pumpkin Tomato Arabidopsis Cottona Cucurbit PTS-1 C-terminal amino acid sequence FLT FLT FLT FLT ISR SEI QEI DIT WTR WTR TRP TRA PPN PPA SYL SYW SYW SYW LDA LDA LDA LAV SHI TRN TRN RAH TGA SGA GAM GAG ASP ASP YNY YNY YNN YDH AAD HIV HIV IYT TNL VNI EMG NLG SIV SIV IVI IVV IVI IVA WDG TEW ADW DAE GDG DRG SAG EEG NSK NAK HHP HHP HYP HYP PSS DIP DTP RLA SVV SIV SEV SVV ALG ALE REL RET KGS INA RAV RHL RVV RPF IAK VAK VAK VAK SKL-C' SKL SRL SRL ARL PRL PRL PRL ARM ARM ARM SRM

Glycolate oxidase

Isocitrate lyase

Hydroxypyruvate reductase Catalase

LPV SKL LPV SKL PTM PSI PSI PNI

SQA DKS CGQ KVA SRL TVK LKA DRS LGQ KLA SRL NVR SQA DKS LGQ KIA SRL NVR SQA DRS LGQ KIA SRL NVR

PTS-2 N'- terminal amino acid sequence AcylCoA oxidase Pumpkin Phalaenopsis Arabidopsis Watermelona Soya bean Rape Cucumber Winter squash Arabidopsis N'-ASPGEPNRTAEDESQAAAR RIERLSLHL MTKEAQMTSLASEHDTQQALR RIQKLSLHL MESRREKNPMTEEESDGLIAAR RIQRLSLHL MQPIPDVNQ MEANSGASD MPHK MQPIPDVNQ MPTDMELSPSNVARH MVFFRSVSAFTRLS RIARISAHL RISRIAGHL RIAMISAHL RIARISAHL RLAVLAAHL RVQGQQSSL TPI LQP SPS HPP RPQ QPS HPP SAA SNS

Malate dehydrogenase

Citrate synthase
a

Indicates there is experimental evidence for the targeting function of this sequence. The bold types indicates the targeting sequence.

the matrix of glyoxysomes during the transitional stage. A variety of proteases have been discovered in leaf peroxisomes but nothing is known about how these might contribute to the selective degradation of enzymes in microbodies (Distefano et al., 1997).

Conclusion
Since the 1980s considerable progress has been made toward understanding the processes that take place in the
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dierent types of microbodies in plant cells and how the proteins that conduct these processes are directed from the cytosol into peroxisomes and glyoxysomes. Yet little is known about how cells maintain and propagate microbodies, how the proteins and lipid molecules of the membrane are assembled, and how proteins pass through the membrane. Nothing is known about how light and levels of metabolites regulate the expression and delivery of proteins to microbodies. Furthermore, there is little knowledge of how metabolites such as fatty acids and carboxylic acids or cofactors such as haem, CoA or/and NAD enter or leave the organelle. Thus, there is much to be

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Plant Cells: Peroxisomes and Glyoxysomes

learned about how microbodies interact with other subcellular molecules and processes.

References
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signal 1 receptor NtPEX5. Proceedings of the National Academy of Sciences of the USA 95(22): 1333613341. Lee MS, Mullen RT and Trelease RN (1997) Oilseed isocitrate lyases lacking their essential type 1 peroxisomal targeting signal are piggybacked to glyoxysomes. Plant Cell 9(2): 185197. Mullen RT, Lee MS, Flynn CR and Trelease RN (1997a) Diverse amino acid residues function within the type 1 peroxisomal targeting signal. Implications for the role of accessory residues upstream of the type 1 peroxisomal targeting signal. Plant Physiology 115(3): 881889. Mullen RT, Lee MS and Trelease RN (1997b) Identication of the peroxisomal targeting signal for cottonseed catalase. Plant Journal 12(2): 313322. Reumann S, Maier E, Heldt HW and Benz R (1998) Permeability properties of the porin of spinach leaf peroxisomes. European Journal of Biochemistry 251(12): 359366. Titus DE and Becker WM (1985) Investigation of the glyoxysome peroxisome transition in germinating cucumber cotyledons using double-label immunoelectron microscopy. Journal of Cell Biology 101: 12891299. Tolbert NE (1981) Metabolic pathways in peroxisomes and glyoxysomes. Annual Review of Biochemistry 50: 133157. Wolins NE and Donaldson RP (1994) Specic binding of the peroxisomal protein targeting sequence to glyoxysomal membranes. Journal of Biological Chemistry 269(2): 11491153. Wolins NE and Donaldson RP (1997) Binding of the peroxisomal targeting sequence SKL is specied by a low-anity site in castor bean glyoxysomal membranes. A domain next to the SKL binds to a highanity site. Plant Physiology 113(3): 943949.

Further Reading
Nishimura M, Hayashi M, Kato A, Yamaguchi K and Mano S (1996) Functional transformation of microbodies in higher plant cells. Cell Structure and Function 21(5): 387393. Olsen LJ and Harada JJ (1995) Peroxisomes and their assembly in higher plants. Annual Review of Plant Physiology and Plant Molecular Biology 46: 123146. Tolbert NE and Essner E (1981) Peroxisomes and glyoxysomes. Journal of Cell Biology 91: 271s283s.

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