Partial structure and properties of the ferredoxin from Rhodymenia palmata

A ferredoxin of MW 11 000 was isolated from the marine alga Rhodymenia palmata (Palmaria palmata). In its oxidised form the ferredoxin had absorption maxima at 276, sh 281, 328, 423 and 465 nm, and contained a single [2Fe-2S] cluster. The midpoint potential of the ferredoxin was ?400 mV and it effectively mediated electron transport in NADP⁺-photoreduction by higher plant chloroplasts, and pyruvate decarboxylation by the phosphoroclastic system of an anacrobic bacterium. The amino acid composition was Lys₃, His₁, Arg₁, Asx₁₂, Thr₉, Ser₈, Glx₁₃, Pro₄, Gly₈, Ala₇, Cys₅, Val₈, Ile₄, Leu₉, Tyr₄, Phe₂; tryptophan and methionine were absent from the molecule. The N-terminal amino acid region consisting of ca half the total amino acid sequence was determined using an automatic sequencer.     

Amino Acid Composition of Cucumber Green Mottle Mosaic Virus Coat Protein and Sequence Determination of Its Tryptic Peptides

Amino acid composition of the CGMMV* coat protein was determined to be as follows: Asp₂₀, Thr₁₀, Ser₂₄, Glu₁₀. Pro₆, Gly₉, Ala₂₁, Val₇, Ile₇, Leu₁₈, Tyr₄, Phe₉, Lys₄, His₁, Arg₈, Trp₂. No terminal α-amino group was detected by dinitrophenylation method. The carboxyl-terminus was found to be serine by hydrazinolysis of the protein and digestion with carboxypeptidase A.

For sequence analysis of the coat protein, tryptic digestion was accomplished at pH 8.0 resulting in ten soluble and several insoluble peptides at pH 4.5. The amino acids contained in soluble peptides accounted for 91 out of 160 residues in the whole protein. The amino acid sequences of ten soluble peptides were determined.

From the similarities of amino acid sequence of the peptides to those of TMV* protein, CGMMV was assumed to be a strain of TMV group.     

Purification,characterisation and amino acid composition of the putative moult-inhibiting hormone (MIH) ofCarcinus maenas (Crustacea,Decapoda)

Summary Using a Y-organ in vitro assay to measure repression of ecdysteroid synthesis in the presence of putative moult-inhibiting hormone (MIH), in conjunction with HPLC separation of sinus gland neuropeptides ofCarcinus maenas, it was found that both the hyperglycemic hormone (CHH) and a novel peptide (argued to represent the MIH) inhibited ecdysteroid synthesis. The latter was purified to homogeneity, and amino acid analysis showed that it is a 61 residue peptide (minimum molecular mass 7,200 Da) with the following amino acid composition: Asx₉; Thr₂; Ser₂; Glx₇; Pro₁; Gly₄; Ala₂; 1/2 Cys₄; Val₄; Met₁; Ile₃; Leu₅; Tyr₁; Phe₃; His₃; Trp₂; Lys₂; Arg₆. The N-terminus appears to be blocked. MIH is at least 20 times more potent than CHH in repressing ecdysteroid synthesis and is active at concentrations of less than 250 pmol/l. There may be structural similarities between CHH and MIH, howeve, MIH displays no CHH radioimmunoreactivity or hyperglycemic activity. The physiological significance of CHH in controlling ecdysteroid titres is not known.Abbreviations CHH hyperglycemic hormone - MIH moult inhibiting hormone - PAGE polyacrylamide gel electrophoresis - RIA radioimmunoassay - SDS sodium dodecyl sulfate - SG smus gland(s) - SGE sinus gland equivalent - TFA trifluoroacetic acid     

The amino acid sequence of human APOA-I, an apolipoprotein isolated from high density lipoproteins.

The complete amino acid sequence of human A-I has been determined by manual and automated Edman degradation of intact and peptide fragments of A-I. A-I is a single chain protein of 243 residues with the following amino acid composition: Asp₁₆, Asn₅, Thr₁₀, Ser₁₅, Glu₂₇, Gln₁₉, Pro₁₀, Gly₁₀, Ala₁₉, Val₁₃, Met₃, Leu₃₇, Tyr₇, Phe₆, Trp₄, Lys₂₁, His₅, and Arg₁₆. The amino acid sequence contains no linear segments of hydrophobic or hydrophilic residues. A detailed correlation of the amino acid sequence, conformation, and self association of A-I will add further insight into the molecular mechanisms involved in protein-protein and protein-lipid interactions.     

Purification and Properties of Trypsin Inhibitor from Hakuhenzu Bean (Dolichos lablab)

A highly purified trypsin inhibitor was obtained from the oriental plant Hakuhenzu bean (Dolichos lablab) by column chromatography on DEAE-Sephadex and gel-filtration on Sephadex G–75. The purified Hakuhenzu bean trypsin inhibitor (HTI) was obtained as a chemically homogeneous protein, and was stable to heat and to enzymes such as pepsin. It shows no obvious maximum at 280 nm in the ultraviolet absorption spectrum, and it contains more than 20% carbohydrate as galactose and 10% hexosamine as glucosamine. The molecular weight of this inhibitor was determined to be approximately 9,500 by gel-filtration. The protein contained 59 residures of amino acids; Lys₃, His₄, Arg₁, Asp₈, Thr₃, Ser₉, Glu₆, Pro₅, Gly₁, Ala₃, l/2Cys₁₀, Val₁, Ile₁, Leu₂, Tyr₁, Phe₁, from which a molecular weight of 6,400 is obtained. No methionine and tryptophan were found in the amino acid composition of the inhibitor. This inhibitor showed inhibitory activity against α-chymotrypsin in addition to trypsin.     

Neutral Proteinases I and II of Aspergillus sojae: Some Physicochemical Properties and Amino Acid Composition

Some physicochemical properties of neutral proteinases I and II, zinc-containing metalloenzymes, from Aspergillus sojae were investigated.

Neutral proteinase I: The enzyme protein had a sedimentation coefficient of 3.90S, an intrinsic viscosity of 0.0315 dl/g, and a partial specific volume, calculated from the amino acid and carbonhydrate composition, of 0.715 cm³/g. The molecular weight was 42,200 from the Yphantis’ procedure, and was 42,500 from the calculation according to the Scheraga-Mandel-kern’s formula. The integral numbers of amino acid residues per molecule calculated on the basis of 42,200 as molecular weight were as follows; Lys₁₆, His₆, Arg₁₃, Trp₈, Asp₅₆, Thr₂₅, Ser₂₃, Glu₃₁, Pro₁₈, Gly₄₀, Ala₃₃, l/2Cys₄, Val₁₁, Met₆, Ile₁₅, Leu₂₅, Tyr₂₀, Р?е₁₀, (amide-ammonia)₂₉, in addition to mannose₆, galactose₁, hexosamine₃.

Neutral proteinase II: The enzyme protein had a sedimentation coefficient of 2.32S, an intrinsic viscosity of 0.0270 dl/g, and a calculated partial specific volume of 0.714 cm³/g. The molecular weight was 16,800 from the Yphantis’ procedure, and was 18,000 from the sedimentation and intrinsic viscosity. The following amino acid compositions was calculated on the basis of 16,800 as molecular weight; Lys₈, His₃, Arg₃, Asp₁₉, Thr₁₇, Ser₁₁, GIu₂₃, Pro₅, Gly₉, Ala₂₄, l/2Cys₄, Val₅, Ile₃, Leu₁₃, Tyr₁₀, Phe₃, (amide-ammonia)₁₅. In the enzyme preparation, neither methionine nor tryptophan was detected and carbohydrate was also absent.

In both neutral proteinases I and II, no free SH group was detected by the PCMB-titration in the presence of 8 M urea.     

Purification of Alkaline Proteinase from Aspergillus Candidus

An alkaline proteinase of Aspergillus Candidus was purified from wheat bran solid culture by batchwise treatment with Amberlite IRC–50 and sequential chromatography on DEAE-cellulose, hydroxylapatite and Sephadex G–100 gel. This purification results in a 18-fold increase of proteolytic activity and the enzyme preparation was homogeneous in sedimentation analysis of the ultracentrifuge and polyacrylamide gel disc electrophoresis. The molecular weight was estimated to be about 23,000 by gel glltration and 22,000 by calculation from the amino acid composition. The enzyme consisted of Lys₁₄, His₄, Arg₃, Asp₂₅, Thr₁₅, Ser₂₃, Glu₁₅, Pro₇, Gly₂₂, Ala₂₄, Met₂, Val₁₆, Ile₁₁, Leu₁₀, Tyr₆, Phe₇, Trp₂ and amide ammonia₁₄ and did not contain cysteine or cystine.     

Photosynthetic Prokaryotes. Cell Differentiation and Functions : Edited by G.C. Papageorgiou and L. Packer Elsevier Biomedical; New York, 1983 405 pages. $65.00

A protein (designated as luffin) with an app. M_r of 26000, which inhibits protein synthesis in rabbit reticulocyte lysate, was purified to homogeneity from the seeds of Luffa cylindria roem by extraction with 20 mM Na phosphate buffer (pH 7.2) containing 0.2 M NaCl, ammonium sulfate fractionation, and chromatography on Sephacryl S-200 and CM-Sephadex C-25. Luffin exhibited 10-times as strong inhibitory activity against protein synthesis in rabbit reticulocyte lysate (IC₅₀, 0.42 ng/ml) as that of ricin A-chain, but it showed only a weak cytotoxicity against murine leukemia L1210 cells, an activity of 1/10⁶ to 1/10⁵ that of ricin.     

Cloning, purification and characterization of two components of phenol hydroxylase from Rhodococcus erythropolis UPV-1

Phenol hydroxylase that catalyzes the conversion of phenol to catechol in Rhodococcus erythropolis UPV-1 was identified as a two-component flavin-dependent monooxygenase. The two proteins are encoded by the genes pheA1 and pheA2, located very closely in the genome. The sequenced pheA1 gene was composed of 1,629 bp encoding a protein of 542 amino acids, whereas the pheA2 gene consisted of 570 bp encoding a protein of 189 amino acids. The deduced amino acid sequences of both genes showed high homology with several two-component aromatic hydroxylases. The genes were cloned separately in cells of Escherichia coli M15 as hexahistidine-tagged proteins, and the recombinant proteins His₆PheA1 and His₆PheA2 were purified and its catalytic activity characterized. His₆PheA1 exists as a homotetramer of four identical subunits of 62 kDa that has no phenol hydroxylase activity on its own. His₆PheA2 is a homodimeric flavin reductase, consisting of two identical subunits of 22 kDa, that uses NAD(P)H in order to reduce flavin adenine dinucleotide (FAD), according to a random sequential kinetic mechanism. The reductase activity was strongly inhibited by thiol-blocking reagents. The hydroxylation of phenol in vitro requires the presence of both His₆PheA1 and His₆PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction.     

Properties of a small basic Peptide from pumpkin seeds

A small basic peptide with an unusual amino acid composition has been isolated from the seeds of pumpkin, Cucurbita maxima. Amino acid analysis and sequence data show the protein to be about 36 residues in length, with an approximate composition Lys₁, Arg₁₄, Asp₃, (Glu + Gln)₁₅, Gly₁, Pro₁, Trp₁. On the basis of composition, the molecular weight is approximately 5000 daltons and the nitrogen content by weight is 20.4%. Twelve amino acids are entirely lacking. The peptide is slightly toxic to mouse B-16 melanoma cells, but its in vivo function is unknown. It does not appear to be derived from cucurbitin, the pumpkin storage globulin; however, it could be a storage peptide involved in nitrogen mobilization during the early stages of germination.     

Analytical studies on crotamine hydrochloride.

The neurotoxin crotamine, which is a basic low molecular weight protein, was isolated, in the form of its hydrochloride, from a South Brazilian rattlesnake (Crotalus durissus crotaminicus, Crotalus durissus terrificus) venom.Disc electrophoresis, carried out in 7% acrylamide by the method of Reisfeld, at pH 4.5, showed a single band for the purified toxin.The toxin showed the following amino acid composition: Asx₂, Ser₃, Glx₂, Pro₃, Gly₅, Cys₅, Met₁, Ileu₁, Leu₁, Tyr₁, Phe₂, Trp₂, Lys₉, His₂, and Arg₂, which corresponds to a minimum molecular weight of 4760. This assignment of minimum molecular weight is supported by the recovery of 1.0 mole of N-terminal $\text{Tyr}\text{5800}\text{g}$ of crotamine by the Sanger dinitrophenol (DNP) method, 1.0 mole of $\text{Tyr}\text{4880}\text{g}$ by the Udenfriend method, and by uv analysis, which gave a value of 4820. The odd number of half-cystine residues ($\text{5}\text{4760}\text{g}$) cannot be explained on the basis of available analytical data.Tyrosine was the only amino-terminal residue detectable by the Sanger DNP method. Glycine was identified as the only carboxyl-terminal residue by the hydrazinolysis method of Akabori and by release upon treatment with yeast carboxypeptidase.The H⁺ electrometric and Cl^? complexometric titrations of crotamine hydrochloride showed that about 13 moles of HCl are bound per 4760 g of the free base.     

Purification and amino acid composition of the hyperglycemic neurohormone from the sinus gland ofOrconectes limosus and comparison with the hormone fromCarcinus maenas

Summary The hyperglycemic neuropeptide was purified to homogeneity from sinus glands of the crayfish,Orconectes limosus, by a simple two-step procedure consisting of preparative polyacrylamide gel electrophoresis followed by gel chromatography on Sephadex G-50. A total of 40 g was obtained from 1,670 lyophilized sinus glands. The molecule consists of 58 amino acid residues plus an undetermined number of Trp residues. The following amino acid composition was found: Asx₈; Thr₂; Ser₃; Glx₆; Pro₁; Gly₂; Ala₃; 1/2 Cys₄; Val₅; Met₁; Ile₃; Leu₅; Tyr₅; Phe₃; Trp_n.d.; His₀; Lys₃; Arg₄. A minimal MW of 6,812 was calculated. The N-terminal residue appears to be blocked. Analysis of the pure hormone on an isoelectric focusing gel gave an pI of 5.04–5.10. The hormone differs from that ofCarcinus (Keller and Wunderer, 1978) in its amino acid composition and pI (Carcinus: 4.55–4.63). The following features are common for both theOrconectes and theCarcinus hormone: the size of the molecule, the blocked N-terminus, the presence of two intrachain disulfide bridges and the general acidic nature. A dose ofOrconectes hormone which is highly effective in this species has no or only very little effect inUca, whereas the same dose ofCarcinus hormone, which is markedly hyperglycemic in the brachyuranUca, exhibits no effect in the astacuranOrconectes. In animals of 10 g live weight, injection of 1.9 pmol/animal causes a threefold increase in the hemolymph glucose level.Dedicated to Professor L.H. Kleinholz on the occasion of his 70^th birthday     

Studies on γ Globulin of Rice Embryo

The molecular weight of γ₃ globulin was determined to be 120,000 daltons by means of both sedimentation equilibrium and gel filtration methods. The protein was composed of 3 identical major and 1 minor subunits, and the molecular weights of them were found to be 35,000 and 13,000 daltons, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major subunit has an arginyl residue as the amino terminal amino acid. The amino acid and carbohydrate composition of γ₃ globulin was determined as follows: Lys₃₇His₃₇Arg₉₂Asp₆₀Glu₁₃₉Gly₁₂₀Ala₈₃Val₇₆Leu₇₀Ile₃₁Pro₅₄Ser₇₇Thr₃₃Cys₁₁Met₉Phe₅₁Tyr₂₆Trp₈ (Amide NH₃)₆₉Hexose₁₅Pentose₄Hexosamine₄. The structure of γ₃ globulin was discussed with comparing that of γ₁ globulin.     

Homology between ricin and Ricinus communie agglutinin: Amino terminal sequence analysis and protein synthesis inhibition studies

The existence of three forms of ricin and two forms of the Ricinus communis agglutinin (RCA) was established using cation exchange chromatography, isoelectric focusing, and polyacrylamide gel electrophoresis. The preparation of the RCA we obtained was 60–75 times more potent than ricin in the agglutination of erythrocytes, but was about 4% as effective as an inhibitor of cell-free protein synthesis. When reduced with 2-mercaptoethanol, the RCA was activated 3000-fold as an inhibitor of cell-free protein synthesis, whereas ricin was activated about 600-fold by the same treatment. A mixture of the RCA A chains was about one-fifth as effective as the ricin A chain in the inhibition of cell-free protein synthesis. The purified polypeptide subunits of the castor bean lectins were subjected to automated Edman degradation. The sequence for 17 of the first 19 residues of the agglutinin A chain was determined. The first seven residues of the ricin A chain were determined and they are identical with those of the RCA A chain. Nineteen turns of Edman degradation on the RCA B chain resulted in the identification of 18 amino acids. The sequence determined for the first 17 residues of the ricin B chain was identical with that of the RCA B chain. It is likely that the identity of the ricin/RCA A and B chain sequences extends further along the polypeptide chains than the sequences we have determined. The similar structural and catalytic potentials of the RCA and ricin suggest that they bear a precursor-product relationship.     

Bothropstoxin-I: Amino acid sequence and function

The complete amino acid sequence of bothropstoxin-I (BthTX-I), a myotoxin isolated fromBothrops jararacussu snake venom, is reported. The results show that BthTX-I is a Lys₄₉ phospholipase A₂ (PLA₂)-like protein composed of a single polypeptide chain of 121 amino acid residues (M _r=13,720), containing one methionine and 14 half-cystines. Although deprived of any detectable PLA₂ activity, BthTX-I reveals a high degree of sequence homology with Asp₄₉-PLA_2s and with other Lys₄₉-myotoxins. Critical mutations—such as Leu₅ for Phe₅; Gln₁₁ for X₁₁; Asn₂₈ for Tyr₂₈; Leu₃₂ for Gly₃₂; Lys₄₉ for Asp₄₉; and Asp₇₁ for Asn₇₁—which are apparently involved with the decreasing or elimination of PLA₂ activity, have been detected. The same mutations occurred in myotoxin II fromBothrops asper venom, but five extra changes—namely, Pro₉₀ for Ser₉₀; Gly₁₁₁ for Asn₁₁₁; His₁₂₀ for Tyr₁₂₀; Phe₁₂₄ for Leu₁₂₄; and Pro₁₃₂ for Ala₁₃₂—have been found relative to myotoxin II.     

Studies on γ Globulin of Rice Embryo

An improved method has been described for the isolation and purification of γ globulin from rice embryo. The method involves the extraction with phosphate buffer, pH 7.0 and ionic strength 0.1, the fractionation in saline solution of ionic strength 0.31, the removal of nucleic acids by precipitation with ammonium sulfate and the gel filtration chromatography on a Sephadex G-200 column. Although the preparations exhibited homogeneous patterns in sedimentation analysis, the electrophoretic patterns on polyacrylamide gels at pH 8.35 and ionic strength 0.11 exhibited at least two components. Three major components, γ₁, γ₂ and γ₃ globulins, were isolated by ion exchange chromatography on a DEAE Sephadex A-50 column. These components were revealed to be homogeneous in electrophoresis as well as sedimentation. N-Terminal amino acid compositions have also been described.

The molecular weight of γ₁ globulin was determined as 2.0 × 10⁵ by the Archibald method, and the intrinsic viscosity, [η], and the sedimentation coefficient, s_{20, w}^°, were found to be 0.0424 dl/g and 7.26S respectively. These values indicated the large asymmetry of the protein. The protein was composed of 18 residues of hexose, 3 residues of pentose, 6 residues of hexosamine and 1751 residues of amino acids: Lys₅₈, His₄₇, Arg₁₄₈, Asp₁₂₆, Glu₂₇₃, Gly₁₆₁, Ala₁₄₄, Val₁₂₁, Leu₁₀₆, Ile₇₂, Pro₈₃, Ser₁₃₆, Thr₄₈, Hyp₆₈, Cys₁₇, Met₁₆, Tyr₄₄, Trp₈, Phe₇₅ and amide ammonia₁₆₃. The N-terminal amino acid analysis suggested that the protein was composed of ten subunits. The properties and the composition were discussed in comparison with those of the 7S globulin of soybean cotyledon.     

Le système protéolytique de Penicillium roqueforti: II - Purification et propriétés de la protéase acide

An extracellular protease has been isolated from the culture medium of Penicillium roqueforti. The enzyme was purified by precipitation with ammonium sulfate, filtration on Bio-gel P 100 columns and chromatography on D.E.A.E.-cellulose columns The purified preparation was homogenous by gel filtration on Bio-gel P 60 and electrophoretical analysis at pH 9.0 and 5.0.The protease exhibited the properties of an acid protease: the optimum pH was 3.5 for casein or hemoglobin hydrolysis and for bovine trypsinogen activation; at 40°C, the enzyme is most stable in the range of pH 3.5 to 5.5; the optimum temperatures was 50°C.E.D.T.A., D.F.P. and sulfhydryl reagents induced no inhibition.The enzyme exhibited a milk clotting activity that was fifty times weaker that the activity of rennin. Its molecular weight, estimated by gel filtration was 33 400. Amino acid composition is: Lys₁₅, His₂, Arg₁, Trp₅, Asp₃₃, Thr₂₇, Glu₁₆, Pro₁₀, Gly₄₀, Ala₂₅, Cys₂, Val₂₁, Ile₂₀, Leu₂₁, Tyr₁₄, Phe₁₉.The properties of this protease were closely similar to that of P. janthinellum and Aspergillus oryzae.     

Identification of the adenine binding site in the ricin toxin A-chain by fluorescence,CD, and electron spin resonance spectroscopy

CD, electron spin resonance, and fluorescence spectroscopy have been utilized to study the adenine binding site of ricin and its toxic A-subunit. At acidic (4.5) and physiological (7.3) pH, adenine or a spin-labeled analogue of adenine, N⁶-(2,2,6,6-tetramethyl-1-oxypiperidin-4-yl) adenine, alters the near uv CD spectra of the ricin A-chain as well as intact ricin, whereas the far uv CD spectra of all proteins remain unchanged. Electron spin resonance data show that the adenine spin-labeled analogue interacts strongly with the A-chain both at pH 4.5 and 7.3, but no or very weak binding is observed for the intact ricin or the isolated B-chain. The adenine spin label gets highly immobilized (2A_II = 65.5G) by the A-chain. The apparent dissociation constant K_d for the toxic A-chain ligand complex is 1.55 × 10^?4 M and 5.6 × 10^?5 M at pH 7.3 and 4.5, respectively. Fluorescence intensity of ricin A-chain bound 1,8-anilinonaphthalenesulfonic acid (ANS) decreases by ～55% at pH 4.5 with the addition of the spin-labeled analogue of adenine, implying that both the ANS and adenine spin label (ADSL) bind to the hydrophobic domain of the A-chain. Fluorescence of the only intrinsic tryptophan probe of the A-chain is also efficiently quenched by ADSL, indicating that the tryptophan residue and the hydrophobic adenine binding site are closely located. All spectroscopic measurements indicate that adenine or its spin-labeled analogue has a single binding site adjacent to the TRP211 residue in the A-chain. Expansion of the A-chain globule and subsequent exposure of the hydrophobic binding site seem to be responsible for the increased binding of adenine at pH 4.5. © 1993 John Wiley & Sons, Inc.     

Ricin A-Chain and Ricin A-Chain Immunotoxins Rapidly Damage Human Endothelial Cells: Implications for Vascular Leak Syndrome

The results of Phase I/II clinical trials indicate that ricin A-chain-containing immunotoxins cause vascular leak syndrome, characterized by hypoalbuminemia with resultant weight gain and edema. Vascular leak syndrome may be a dose-limiting factor during treatment with ricin A-chain-containing immunotoxins. In this report, we determined the effect of ricin A-chain and ricin A-chain-containing immunotoxins on human umbilical vein endothelial cells with the aim of developing an in vitro model to study vascular leak syndrome. The major findings of our study are: (1) Human umbilical vein endothelial cells undergo rapid and dramatic changes in morphology after treatment with ricin A-chain and ricin A-chain-containing immunotoxins. These changes include rounding of the cells and, eventually, the formation of gaps between them. (2) The permeability of human umbilical vein endothelial cell monolayers to passage of molecules increases after exposure to ricin A-chain or ricin A-chain-containing immunotoxins and this is consistent with the morphologic changes. (3) Human umbilical vein endothelial cells bind ¹²⁵ I-rRTA in a dose-dependent manner but binding is not specific. (4) Human umbilical vein endothelial cells are moderately more sensitive to ricin A-chain-induced inhibition of protein synthesis and proliferation than simian virus-transformed mouse endothelial cells. (5) The morphologic changes are observed 1 h after exposure to the toxins, whereas inhibition of protein synthesis is not detectable until 4 h after a similar exposure. The in vitro model represents a first step in dissecting the complex events which occur in cancer patients who develop vascular leak syndrome after treatment with ricin A-chain-containing immunotoxins.