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Evidence for anion-translocating plant uncoupling mitochondrial protein in potato mitochondria.

Transport properties of plant mitochondria from potato tubers were investigated using the swelling technique and membrane potential measurements. Proton-dependent swelling of fatty acid-depleted mitochondria in potassium acetate with valinomycin was possible only in the presence of fatty acids (linoleic acid and 12-(4-azido-2-nitrophenylamino)dodecanoic acid) and was inhibited by various purine nucleotides including ATP, GDP, and GTP. Swelling representing uptake of hexanesulfonate was also inhibited by purine nucleotides. Also, the membrane potential of fatty acid-depleted potato mitochondria energized by succinate declined upon the addition of linoleic acid or 12-(4-azido-2-nitrophenylamino)dodecanoic acid, and this decrease was prevented by ATP and other purine nucleotides. These transport activities are identical to those reported for brown adipose tissue mitochondria and related to the uncoupling protein; therefore, we ascribed them to the plant mitochondrial uncoupling protein (PUMP). A major difference between plant and mammalian uncoupling protein is that PUMP transports small hydrophilic anions such as Cl- very slowly, if at all. We suggest that PUMP may play an important role in plant physiology, where a regulated uncoupling and thermogenesis can proceed during fruit and seed development.

Alkanesulfonates↗

A conserved domain of the viviparous-1 gene product enhances the DNA binding activity of the bZIP protein EmBP-1 and other transcription factors.

The maize VP1 protein is a seed-specific regulator of gene expression that effects the expression of a subset of abscisic acid (ABA)-regulated genes that are expressed during the maturation program of the seed. In addition, VP1 has pleiotropic effects on seed development that are not related to ABA. In transient expression assays, VP1 has been shown to transactivate gene expression through at least two distinct promoter elements: the G boxes from the ABA-inducible wheat Em gene and the SphI box from the maize C1 gene. We have investigated how VP1 can transactivate gene expression through diverse promoter elements by analyzing its association in vitro with EmBP-1, a factor that binds the Em promoter. We demonstrate that VP1 can greatly enhance the DNA binding activity of EmBP-1 in a gel retardation assay. This enhancing activity has also been observed on transcription factors as diverse as Opaque-2, Max, Sp1, and NF-kappaB. Deletion of a small but highly conserved region (BR2) in VP1 eliminates the enhancement in vitro as well as the ability of VP1 to transactivate Em gene expression in a transient expression assay. A 40-amino acid fragment from VP1 sandwiched between the maltose-binding protein and LacZ can confer the enhancement function to this fusion protein in vitro. A weak and relatively nonspecific interaction between BR2 and DNA is demonstrated by UV cross-linking. The in vitro properties we observe for VP1 might explain the regulatory effects of VP1 on a diverse set of genes and why mutations in the vp1 locus have pleiotropic effects.

Base Sequence↗

Role of individual disulfide bonds in the structural maturation of a low molecular weight glutenin subunit.

Gliadins and glutenins are the major storage proteins that accumulate in wheat endosperm cells during seed development. Although gliadins are mainly monomeric, glutenins consist of very large disulfide-linked polymers made up of high molecular weight and low molecular weight subunits. These polymers are among the largest protein molecules known in nature and are the most important determinants of the viscoelastic properties of gluten. As a first step toward the elucidation of the folding and assembly pathways that lead to glutenin polymer formation, we have exploited an in vitro system composed of wheat germ extract and bean microsomes to examine the role of disulfide bonds in the structural maturation of a low molecular weight glutenin subunit. When conditions allowing the formation of disulfide bonds were established, the in vitro synthesized low molecular weight glutenin subunit was recovered in monomeric form containing intrachain disulfide bonds. Conversely, synthesis under conditions that did not favor the formation of disulfide bonds led to the production of large aggregates from which the polypeptides could not be rescued by the post-translational generation of a more oxidizing environment. These results indicate that disulfide bond formation is essential for the conformational maturation of the low molecular weight glutenin subunit and suggest that early folding steps may play an important role in this process, allowing the timely pairing of critical cysteine residues. To determine which cysteines were important to maintain the protein in monomeric form, we prepared a set of mutants containing selected cysteine to serine substitutions. Our results show that two conserved cysteine residues form a critical disulfide bond that is essential in preventing the exposure of adhesive domains and the consequent formation of aberrant aggregates.

Base Sequence↗

A multifunctional acyl-acyl carrier protein desaturase from Hedera helix L. (English ivy) can synthesize 16- and 18-carbon monoene and diene products.

A desaturase with 83% sequence identity to the coriander delta(4)-16:0-ACP desaturase was isolated from developing seeds of Hedera helix (English ivy). Expression of the ivy desaturase in Arabidopsis resulted in the accumulation of 16:1delta(4) and its expected elongation product 18:1delta(6) (petroselinic acid). Expression in Escherichia coli resulted in the accumulation of soluble, active protein that was purified to apparent homogeneity. In vitro assays confirmed delta(4) desaturation with 16:0-ACP; however, with 18:0-acyl acyl carrier protein (ACP) desaturation occurred at the delta(9) position. The ivy desaturase also converted 16:1delta(9)-ACP and 18:1delta(9)-ACP to the corresponding delta(4,9) dienes. These data suggest at least two distinct substrate binding modes, one placing C4 at the diiron active site and the other placing C9 at the active site. In the latter case, 18:0 would likely bind in an extended conformation as described for the castor desaturase with 9-carbons accommodated in the cavity beyond the dirron site. However, delta(4) desaturation would require the accommodation of 12 carbons for C16 substrates or 14 carbons for C18 substrates. The amino acids lining the substrate binding cavity of ivy and castor desaturases are conserved except for T117R and P179I (castor/ivy). Paradoxically, both substitutions, when introduced into the castor desaturase, favored the binding of shorter acyl chains. Thus, it seems likely that delta(4) desaturation would require a non-extended, perhaps U-shaped, substrate conformation. A cis double bond may facilitate the initiation of such a non-extended conformation in the monounsaturated substrates. The multifunctional properties of the ivy desaturase make it well suited for further dissection of the determinants of regiospecificity.

Amino Acid Sequence↗

Cloning and characterization of two novel lipid transfer protein I genes in Vigna radiata.

Two full-length lipid transfer protein (LTP) cDNAs were isolated from mungbean (Vigna radiata) and designated Vrltp1 and Vrltp2. The deduced amino acid sequences contain the two highly conserved pentapeptides characteristic of plant LTPIs suggesting these Vrltps belong to the LTPI gene family. Vrltp1 mRNA was detected in developing seeds, but Vrltp2 mRNA was not. Within the vegetative tissues, the Vrltp1 and Vrltp2 mRNAs were present only in leaves and stems, but not root tips. Salt and dehydration stresses and exogenous abscisic acid (ABA) treatments resulted in increased mRNA levels of both Vrltps in leaves. We suggest that these unique Vrltps are specific to growing shoot tissues, and may play an important role in plant acclimation to water stress.

Amino Acid Sequence↗

Isolation and characterization of TaDof1 transcription factor in wheat (Triticum. aestivum. L).

The Dof (DNA binding with one finger) proteins are plant specific transcription factors. Dof proteins are apparently encoded by a multiple gene family in higher plants. However, only one Dof gene, WPBF, was reported in wheat. In this study, a member of Dof gene family, TaDof1, was cloned from wheat. TaDof1 encode 291 amino acids, with a predicted molecular mass of 30.348 kDa. At its N-terminal end, a 52 amino acid stretch typical of Dof domain and two serine-rich stretches were observed. Sequence alignment indicated that, in Dof domain, TaDof1 share more than 75% identity with other Dof proteins of different species. TaDof1 was expressed highly in leaves and sheaths, but lowly in roots, and constitutively expressed in developing seeds of 2-12 DAP. It was interesting to note that TaDof1 was differentially expressed between hybrids F1 and parents in root, sheath and leaf. The implication of the differential expression patterns of TaDof1 was discussed in related to the up-regulation of C4 pathway related gene in hybrid rice and heterosis.

Amino Acid Sequence↗

Transient N-acetylglucosamine in the biosynthesis of phytohemagglutinin: attachment in the Golgi apparatus and removal in protein bodies.

Cotyledons of the common bean (Phaseolus vulgaris L.) synthesize large amounts of the lectin phytohemagglutinin (PHA) during seed development. The polypeptides of PHA are synthesized by endoplasmic reticulum-bound polysomes and co-translationally glycosylated, pass through the Golgi complex, and accumulate in protein bodies, which constitute the lysosomal compartment in these cells. Some of the high-mannose sidechains of PHA are modified in the Golgi complex, and in mature PHA they contain N-acetylglucosamine, mannose, fucose, and xylose in the molar ratios 2, 3.8, 0.6, and 0.5. The results reported here show that the Golgi complex is also the site of additional N-acetylglucosamine incorporation into the modified sidechains. When developing cotyledons are labeled with [3H]glucosamine and glycopeptides of PHA present in the Golgi complex isolated, the radioactivity can be released as [3H]N-acetylglucosamine by digestion of the glycopeptides with beta-N-acetylglucosaminidase, indicating that the residues are in a terminal position. Arrival of PHA in the protein bodies is followed by the slow removal of these terminal N-acetylglucosamine residues, resulting in a decrease in the Mr of the modified sidechains. The biosynthetic intermediates of the glycoproteins destined for the lysosomal compartments of animal cells contain high-mannose sidechains modified by phosphate groups covered by N-acetylglucosamine that is labile to mild acid treatment. When cotyledons are labeled with [32P]orthophosphate, there is no radioactivity in PHA obtained from any of the subcellular fractions. There is also no release of radioactivity when [3H]glucosamine-labeled glycopeptides obtained from PHA in the Golgi complex are subjected to mild acid hydrolysis. These results indicate that the sorting-signals and posttranslational processing steps for proteins that are transported to the lysosomal compartment are different in plant cells and animal cells.

Acetylglucosamine↗

The SHOOTLESS2 and SHOOTLESS1 genes are involved in both initiation and maintenance of the shoot apical meristem through regulating the number of indeterminate cells.

To characterize the SHL2 and SHL1 genes in detail, we analyzed three strains carrying weak alleles of SHL2, shl2-6, shl2-7, and shl2-8, and one weak allele of SHL1, shl1-3. In contrast to strong alleles, which result in lack of shoot meristem, strains bearing these weak alleles formed shoot meristem frequently during embryogenesis. In shl2-6 and shl2-7 mutants, the meristem was lost during seed development. Only the shl2-8 mutant could survive after germination, but it showed abnormal initiation pattern and morphology of leaves. In strains bearing the weak alleles, the shoot meristem was composed of a small number of indeterminate cells and ultimately converted into leaf primordium. The shl1-3 mutant showed phenotypes similar to those of shl2-8. Thus SHL2 and SHL1 are required for both initiation and maintenance of shoot meristem. In shl2 mutants, there was a positive correlation between the size of the expression domain of OSH1 representing the number of indeterminate cells, the frequency of shoot meristem initiation, and the duration of meristem survival. Thus the shoot meristem will not initiate in an "all-or-nothing" fashion, but is formed in various degrees depending on the strength of the alleles. Double-mutant analyses indicate that SHL2 functions upstream of SHO to establish proper organization of the shoot meristem.

Homeodomain Proteins↗

B cell development and activation defects resulting in xid-like immunodeficiency in BLNK/SLP-65-deficient mice.

Engagement of the B cell receptor (BCR) leads to the activation of tyrosine kinases and other signaling molecules that ultimately determine the type and magnitude of the B lymphocyte's cellular response. The adaptor protein BLNK/SLP-65 plays a pivotal role in BCR signal transduction by coupling Syk activation to downstream elements such as Grb2, phospholipase C-gamma, Vav and Nck. We have generated BLNK(-/-) mice to determine the physiological role of this protein in B cell development and activation. BLNK(-/-) mice exhibit an incomplete block in B cell development with a severe inhibition of pro-B to pre-B cell differentiation. BLNK(-/-) sIgM(+) cells can develop, seed the peripheral lymphoid tissues and accumulate in numbers overtime. However, these mutant B cells failed to mature and are non-responsive to BCR cross-linking in terms of proliferation and up-regulation of activation markers such as CD69 and CD86 (B7-2). In addition, the CD5(+) subset of B cells is absent. The immune response to T cell-independent antigen but not T cell-dependent antigen is also impaired. Overall, the phenotype of BLNK(-/-) mice bears a striking resemblance to that of xid mice which is the murine model of human XLA that has a mutation in Bruton's tyrosine kinase. This raises the interesting possibility that mutation in BLNK/SLP-65 may be responsible for certain human immunodeficiencies.

Adaptor Proteins, Signal Transducing↗

The genetic basis of naturally occurring pollen color dimorphisms in Nigella degenii (Ranunculaceae).

Nigella degenii ssp. barbro and ssp. jenny differ from related taxa in being dimorphic for pollen color, with some plants having dark pollen and others light pollen. In this study we performed experimental crosses to determine whether the difference in pollen color is governed by few or many loci and whether the two subspecies utilize the same gene to control pollen color. Patterns of segregation in crosses between morphs show that dark pollen is dominant over light pollen and that a single major gene is responsible for most of the variation in pollen color. Consequently it should be relatively easy for pollen color dimorphisms to establish and spread in these subspecies. Aberrant segregation ratios were attributed to genetic factors that reduced the expression of the allele conferring dark pollen or processes that sorted between color morphs during seed development. Crosses between dark pollen plants from different subspecies showed signs of complementation in the F2 generation, but the frequency of the light morph was too low to support a model involving complementary action of recessive alleles at two separate loci. Based on this and other observations, we hypothesize that the pollen color difference is controlled by the same major locus in the two subspecies.

Analysis of Variance↗

Dietary supplementation with high-selenium soy protein reduces pulmonary metastasis of melanoma cells in mice.

The effect of high-selenium (Se) soy protein on pulmonary metastasis of murine B16BL6 melanoma cells was investigated in male C57BL6 mice. Isolated soy proteins (ISP) from soybeans grown with and without Se foliar application during seed development were compared. Five diets were studied, a basal AIN-93G diet or a basal diet containing 10% low-Se ISP, 5% low-Se + 5% high-Se ISP, 10% high-Se ISP, or 10% low-Se ISP supplemented with Se equivalent to that of the 10% high-Se ISP diet. The Se concentrations of the 5 diets were 0.13, 0.13, 1.9, 3.6, and 3.0 microg/g, respectively. Mice were fed the diet for 2 wk before and 2 wk after an i.v. injection of 5 x 10(4) viable cells. At necropsy, the number and size of tumors that had developed in the lungs were determined. In the control group, 13/18 mice exhibited > or = 50 tumors. The numbers of mice with > or = 50 tumors were 8/18, 7/18, 3/18, and 6/17 in the ISP-fed groups, respectively. The differences between the 10% high-Se ISP group, the Se-supplemented 10% low-Se group, and the control were significant (P < 0.05). Dietary supplementation with 10% low-Se ISP significantly decreased the mean number of tumors per group and the tumor size compared with the control. A greater reduction in these variables occurred in mice fed the 10% high-Se ISP diet. The inhibition by the Se-supplemented 10% low-Se ISP diet was similar to that by the 10% high-Se ISP diet. The whole-blood Se concentration was inversely related to the tumor number (R = -0.87, P = 0.052), tumor cross-sectional area (R = -0.91, P < 0.05), and tumor volume (R = -0.93, P < 0.05). These findings suggest that Se is responsible for the greater antimetastatic effect of the high-Se ISP. We conclude that the high-Se soy protein has a greater inhibitory effect than the low-Se soy protein on pulmonary metastasis of melanoma cells in mice.

Animals↗

Roles of cell-wall invertases and monosaccharide transporters in the growth and development of Arabidopsis.

The hydrolysis of sucrose by cell-wall invertases (cwINV) and the subsequent import of hexoses into target cells appears to be crucial for appropriate metabolism, growth and differentiation in plants. Hexose uptake from the apoplast is catalysed by monosaccharide/H+ symporters (Sugar Transport Proteins or STPs), which have the potential to sense sugars. Import of extracellular hexoses may generate signals to orchestrate cellular activities, or simply feed metabolic pathways distinct from those fed by sucrose. It is predicted that Arabidopsis has six cwINV genes and at least 14 STP genes. These genes show different spatial and temporal patterns of expression, and several knock-out mutants have been isolated for analysis. AtSTP1 transports glucose, galactose, xylose, and mannose, but not fructose. It accounts for the majority of the AtSTP activity in vegetative tissues and its activity is markedly repressed by treatment with exogenous sugars. These observations are consistent with a role in the retrieval of cell-wall-derived sugars, for example, during carbohydrate limitation or cell expansion. The AtSTP1 gene is also expressed in developing seeds, where it might be responsible for the uptake of glucose derived from imported sucrose. The large number of AtcwINV and AtSTP genes, together with complex patterns of expression for each, and the possibility that each protein may have more than one physiological function, provides the plant with the potential for a multiplicity of patterns of monosaccharide utilization to direct growth and differentiation or to respond flexibly to changing environmental conditions.

Arabidopsis↗

Phytosulphokine gene regulation during maize (Zea mays L.) reproduction.

The sulphated pentapeptide phytosulphokine (PSK) was identified as a substance that promotes cell division in low-density suspension cultures and has been implicated in various aspects of tissue differentiation in plants. The peptide is derived from PSK precursor proteins that are encoded by small gene families. The physiological roles of PSK are still not clearly defined and little is known about expression of members of the PSK precursor gene family in any plant species. In this study, highly regulated tissue and cell type-specific expression are described for four PSK genes from maize (Zea mays L.) in female and male gametophytes, and during seed development. ZmPSK1 and ZmPSK3 were specifically and differentially expressed in cells of female and male gametophytes and in female and male gametes. In anthers ZmPSK1 or ZmPSK3 transcripts were found, for example, at high levels in secretory tapetal cells which support developing microspores. ZmPSK1 mRNA was abundant in mature pollen including sperm cells. ZmPSK1 and ZmPSK3 transcripts were also detected in egg and central cells of the female gametophyte and ZmPSK1 mRNA was present in synergids, indicating that the PSK peptide probably plays a role during gametogenesis, pollen germination, and fertilization. In developing maize kernels all four ZmPSK genes were expressed, albeit with striking differences in their expression patterns. It is proposed here that PSK is required for numerous but defined processes during gametophyte and early sporophyte development. In general, PSK availability appears to be controlled through transcriptional regulation in a tissue and cell type-specific and development-dependent manner.

Amino Acid Sequence↗

The leader intron of Arabidopsis thaliana genes encoding cytochrome c oxidase subunit 5c promotes high-level expression by increasing transcript abundance and translation efficiency.

The involvement of regions located upstream of the translation start site in the expression of two Arabidopsis thaliana nuclear COX5c genes encoding subunit 5c of mitochondrial cytochrome c oxidase has been analysed. It was observed that these regions, which include a leader intron, direct the tissue-specific expression of the gus reporter gene, mainly in root and shoot meristems, actively growing tissues and vascular strands. Expression was also observed in flowers, specifically localized in anthers, stigma, and the receptacle, and in developing seeds. GUS activity measurements in protein extracts from transformed plants indicated that expression levels are higher than those observed with the constitutive CaMV 35S promoter. Removal of the leader intron produced a significant decrease in expression to values only slightly higher than those observed with a promoterless gus gene. Histochemical staining of plants transformed with the intronless construct revealed expression only in pollen, suggesting that regulatory elements capable of directing pollen-specific expression are present upstream of the intron. The COX5c-2 intron also increased GUS expression levels when fused in the correct orientation with the promoter of the unrelated COX5b-1 gene. Comparison of GUS activity values with the transcript levels suggests that the intron also increases translation efficiency of the corresponding mRNA. The results obtained point to an essential role of the intron present in the 5'-non-coding region of all known COX5c genes in directing the expression of these genes in plants.

5' Untranslated Regions↗

Cell-specific expression of genes of the lipid transfer protein family from Arabidopsis thaliana.

We have characterized three cDNAs from a gene family encoding lipid transfer proteins, LTP, from Arabidopsis thaliana (Wassilewskija). In addition to the already characterized Ltp1, our analysis includes Ltp2 and Ltp3, two sequences previously known as expressed sequence tags (EST) only. The deduced amino acid sequences of the three cDNAs share 56 to 57% identity and show unique tissue- and cell-specific expression. Genes Ltp1 and LTp2 are located within approximately 1.4 kb of each other in tandem orientation. RNA hydridizations showed that all three LTP are expressed in flowering meristems, flowers and developing seeds. Ltp1 is expressed in leaves in addition. Ltp3, though not Ltp2, is also expressed in a short segment of the stem close to the flowering meristem. In contrast to the epidermis-specific Ltp1, both Ltp2 and Ltp3 are not restricted to the epidermis, but are also expressed in sub-epidermal layers of the organs in which they are found. In the upper stem segment, Ltp3 is predominantly cortical. It appears that the expression of these three cDNAs is sufficient to account for the formation of LTP in all meristematic and expanding cells of the aboveground plant. Evolutionary analysis allows the conclusion that each Ltp belongs to a different sub-family of genes. Additionally, parsimony analysis provides evidence that several copies of Ltp genes already existed in ancestors of the Brassicaceae family.

Amino Acid Sequence↗

Characterization of a flower-specific gene encoding a putative myrosinase binding protein in Arabidopsis thaliana.

A cDNA clone, 4B-1, previously isolated by differential screening is preferentially expressed in floral organs of Arabidopsis thaliana. Characterization of the full length cDNA and the genetic locus corresponding to 4B-1 cDNA revealed that it potentially encodes a myrosinase binding protein (MBP) which is presumably present in a large myrosinase complex. The deduced amino acid sequence of the polypeptide encoded by cDNA clone (designated f-AtMBP) appeared to consist of two parts: one region at the C-terminal half representing overall homology with AtMBP, an MBP homologue in A. thaliana, and the other at an extended N-terminal region of about 150 amino acids showing significant identity with the N-terminal region of the MBP-related protein reported in Brassica. Expression analysis by RNA blot and in situ hybridization showed that f-AtMBP was specifically expressed in floral meristems, pistils, stamens, petals, and ovules of immature flowers, but no expression was observed in the specialized cells called the myrosin cells in the hypocotyl and cotyledons of developing seeds where myrosinase enzymes are normally found. Although MBPs and MBP-related proteins are considered to be inducible by exogenous application of signal molecules and physical wounding, we found that f-AtMBP expression was not activated by such treatment, suggesting that f-AtMBP is a novel type of MBP specific to floral organs.

Amino Acid Sequence↗

Accumulation of a fusion protein containing 2S albumin induces novel vesicles in vegetative cells of Arabidopsis.

We have previously reported that precursor-accumulating (PAC) vesicles found exclusively in developing seeds are involved in a transport of seed storage proteins, such as 2S albumin, from the endoplasmic reticulum to protein-storage vacuoles. Here, we constructed chimeric genes that encode fusion proteins consisting of both various lengths of polypeptides derived from pumpkin 2S albumin and a selectable marker enzyme, phosphinothricin acetyltransferase. The chimeric genes were expressed in transgenic Arabidopsis in order to investigate the mechanism of the PAC vesicle formation. A fusion protein expressed by one of the chimeric genes is accumulated as a proprotein-precursor form, and localized in novel vesicles of vegetative cells. The vesicles show distinct features that well much to the PAC vesicles. Despite of the accumulation of the fusion protein, the transgenic Arabidopsis is still sensitive to phosphinothricin. Phosphinothricin acetyltransferase contained in the fusion protein is obviously compartmentalized in the PAC-like vesicles that do not permit the detoxification of this herbicide. These results indicate that the PAC-like vesicle can be induced in vegetative cells by the ectopic expression of the protein that is destined to be compartmentalized into the PAC vesicles.

Acetyltransferases↗

Purification and characterization of phospholipase D (PLD) from rice (Oryza sativa L.) and cloning of cDNA for PLD from rice and maize (Zea mays L.).

Phospholipase D (PLD) was purified to high homogeneity from rice bran (Oryza sativa L.). Two peaks of PLD activity were resolved by Mono Q anion-exchange chromatography. The molecular mass of PLD in both peaks was 82 kDa on SDS-PAGE and 78 kDa in gel filtration. Antibodies raised against the protein in one of the peaks precipitated the enzyme activities in both peaks. Enzymatic characteristics of PLD in the two peaks were identical except for a difference of 0.1 in the isoelectric points. Sequence analysis covering more than 10% of the amino acids of the proteins and peptide mapping did not detect any difference in the primary structure of the proteins. A cDNA for PLD was isolated from rice and it encoded a protein of 812 residues. The N-terminal sequences of purified PLDs matched the deduced amino acid sequence starting from residue 47. A Northern blot showed this gene was expressed in leaves, roots, developing seeds and cultured cells, and a Southern blot detected a single band of rice genomic DNA hybridizing to the cDNA. A cDNA for PLD was also isolated from maize. The similarity of the deduced amino acid sequences of PLD was 90% between rice and maize, 73% between the cereals and castor bean.

Amino Acid Sequence↗