Medical Forum / Diseases and Disorders / Herpes / November 2005
L-Canavanine
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Perl Molson - 31 Oct 2005 15:21 GMT What is it?
L-Canavanine is one of over 300 documented non-protein amino acids, also known as secondary metabolites or anti-metabolites, which exist in plants (most commonly in legumes and their seeds). Essentially, they assist in the breakdown of the 20 known amino acids common in both plants and animals. L-Canavanine's chemical structure is quite complex and defined as follows:
H2N-C(=NH)-NH-O-CH2-CH2-CH2-CH(NH2)CO2H
Specifically, L-Canavanine is an L-arginine antimetabolite. In the Sutherlandia plant, L-Canavanine is present at levels of just over 2.0 mg per dry leaf gram. It acts as an anti-predatory agent in the plant, protecting it against various insects.
L-Canavanine has been shown to be an inhibitor (selective) of inducible nitric oxide synthase (iNOS), with possible usage in the treatment of alcohol induced brain damage and certain other neurological disorders.
What are its benefits?
L-Canavanine has been identified by G. A Rosenthal, of the Biochemical Ecology Laboratory at The University of Kentucky, as a likely therapeutic agent for human pancreatic and some other cancer types. It has been demonstrated by N. Suzuki, A. Sakomoto and R. Ogawa as having strong potential for the treatment of septic shock and chronic inflammation. These are just a few of numerous research papers on the substance.
There are a number of patents associated with L-Canavanine which indicate it's applicability in the treatment in a number of ailments. Two of the main examples are: Crooks, Rosenthal and Gerald were granted a patent in 1994 for the use of the substance to treat pancreatic cancer (Patent No. 5,552,440) and Karson, Lyon and Freeman being granted a patent that uses the substance in the treatment of numerous brain disorders in 1996 (Patent No. 5,527,825). In patent No. 6,346,519, E. Petrus gives L-Canavanine a partial role in the treatment of arthritis.
More research is currently underway on the substance and its applicability to certain other conditions. Details will be placed here as soon as confirmed.
Is a patent total proof that L-Canavanine is effective?
The tests, experiments and theory associated with the above have met a defined standard of proof with the US Patent office, as evidenced by the awarding of the patents. Despite this, we do not regard the awarding of the patents as demonstrable evidence for the efficacy of L-Canavanine in themselves. But critically, the above findings do correlate with the widespread anecdotal evidence we have taken from individuals who have used Sutherlandia and the fact that their ancestors, going back hundreds of years, have also used the plants for the same purpose, with very positive results.
Are there any side effects?
If taken in severe overdose quantities (i.e. 1,000 mg per day for 12 months), L-Canavanine can cause thinning of the blood. Furthermore, it can exacerbate the symptoms of Lupus, a disease where the body's own immune system starts to attack various organs. Individuals with kidney conditions must consult their physician before taking this substance.
At recommended dosage levels, the L-Canavanine in Sutherlandia Frutescens Tablets has not been shown to cause side effects.
Perl Molson - 31 Oct 2005 15:41 GMT Effects of canavanine treatment on herpesvirus morphogenesis in cultured cells.
Smith JD, Moore DM.
L-Canavanine, a naturally occurring analog of arginine, effectively inhibited the morphogenesis of herpes simplex virus (HSV) and human cytomegalovirus (HCMV) when added at the time of infection, but allowed the expression of several cytopathic changes. Exposure to canavanine at progressively later times ultimately led to qualitatively normal virion maturation. Under no conditions were morphologically aberrant viral particles observed. However, HSV-infected cells treated at 3 h postinfection or later did contain distinctive cytoplasmic inclusions resembling HCMV dense bodies. Recovery experiments showed that HCMV-infected cells exposed to canavanine for 1-5 days could support normal viral morphogenesis when washed free of this agent.
PMID: 6282774 [PubMed - indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstra ct&list_uids=6282774&query_hl=2
Perl Molson - 31 Oct 2005 16:06 GMT Specificity in protein biosynthesis
The general problem of recognition of their substrates by the aminoacyl-tRNA synthetases has been the subject of intensive research for several decades and, with the advent of molecular biological techniques together with available X-ray crystallographic structures, considerable progress towards the understanding of tRNA recognition has been achieved. However, investigations into the recognition of the amino acid and the mechanisms involved in guaranteeing accuracy during aminoacylation have, with the tools of recombinant DNA, only recently been reinitiated, following detailed biochemical studies in the mid-'70s. These defined the pathways of "chemical proofreading" or "hydrolytic editing" by which a transiently activated "wrong" amino acid is corrected by the enzyme itself, in a proofreading step.
Renewed interest in recent years, with the possibilities available through the methods of genetics and molecular biology, boosted the hope that attempts at solving this central enzymological puzzle may be able to pinpoint regions or even residues which are essential for maintaining fidelity. Mutational analysis established conserved residues that are important for proofreading. However, it is quite possible, based on phenomenological observations, that several different mechanisms may have evolved.
It is reasonable then, to approach the elucidation of this/these mechanism/s from an alternative angle; using the systems provided by Nature, in the form of inhibitory non-protein amino acids. A comparison between the enzyme from a non-producer, from a producer and from a resistant seed predator (using the possibilities offered by recombinant techniques to obtain well-defined, purified components) will give important information on how nature has solved the problem of not incorporating these non-protein amino acids into the growing protein chain. Of the thousands of known non-protein amino acids (NPA), about 300 occur in plants. They are found mostly in a small number of families, such as the Leguminosae, Cucurbitaceae, Sapindacae, Aceraceae and Hippocastenaceae. Many of these NPA are structurally similar to the components of common proteins. The incorporation of NPAs into proteins may be associated with autoimmune diseases in humans. Furthermore, there is evidence of a phenotypic conversion of ras-transformed human cells to normal due to incorporation of the tyrosine analogue, azatyrosine, into cellular protein. One NPA that has received some attention is canavanine, (L-2-amino-4-(guanidinooxy)butyric acid), the guanidinooxy structural analogue of arginine (Fig. 1).
Fig. 1: L-Arginine (top), L-Canavanine (bottom).
Since its discovery in the 1920's in the seeds of jack bean (Canavalia ensiformis), massive accumulation of this toxic secondary metabolite has been established in the seeds of at least 1500 members of the Lotoideae; a major subdivision of the Fabaceae. Canavanine storage in the seeds of such plants can account for more than 10% of the seed dry matter. The preponderance of seed canavanine, its depletion during growth of the seedlings, its extreme toxicity to actual (insects, rodents) and serendipitous (humans) seed predators, and its high nitrogen content all point to the conclusion that the plant has constructed a compound that is simultaneously a storage compound and a major seed defence. This strategy is commonplace in legume seeds with a variety of secondary compounds. The evolutionary history is undoubtedly that of evolution of toxicity in compounds that were initially purely storage in function; a change with obvious adaptive value. The toxicity of canavanine to a non-producer species ranging from bacteria to insects, to plants and mammalian cells has been extensively described. Canavanine binds to nearly every enzyme that utilizes arginine. The mechanism of its toxicity has, however, been localized to its incorporation into proteins giving rise to functionally aberrant polypeptides. The loss or reduction in activity in canavanine-containing proteins has been observed for antibacterial proteins and for lysozyme into which canavanine had been randomly incorporated. Canavanine containing proteins are also preferentially degraded, in vivo. Allende and Allende in the '60s first documented the attachment of canavanine to tRNAArg by the rat liver arginyl-tRNA synthetase.
This lack of discrimination by an enzyme belonging to a family of enzymes whose accuracy in recognizing their substrate is essential in maintaining an effective protein synthesizing machinery, was confirmed for the enzyme from E.coli and Neurospora while an assay with a crude extract from the producer, Canavalia, showed a much reduced activity.
The accumulation of canavanine in producers provides nitrogen storage for the growing seedling and also provides an effective protective chemical barrier to predation and disease and raises the question as to how the producer itself avoids autotoxicity. In addition to the need for a detoxifying mechanism for the producer being self-evident, there are also organisms that are themselves non-producers but have specialized in feeding on the producer. Such a situation has been found in the bruchid beetle Caryedes brasiliensis whose larvae infest and feed on Dioclea violacea seeds containing up to 13% canavanine.
In this project, we propose to isolate, clone and overexpress the gene(s) for arginyl-tRNA synthetase from at least two canavanine producers (Canavalia ensiformis, Dioclea violaceae (formerly, megacarpa)), a closely related non producer (Glycin max) and a resistant seed predator (Caryedes brasiliensis). We will study the structure/function of the gene products in order to identify the molecular mechanism(s) involved in this example of allelochemical detoxification while establishing the discriminatory elements involved in the recognition of arginine/canavanine by arginyl-tRNA synthetase. Additionally, should the adult Caryedes prove to be canavanine sensitive, by studying the argS genes/regulatory elements/gene products we intend to obtain information on the developmental regulation that occurs during insect metamorphosis generating a canavanine-resistant (Caryedes larvae) or non-resistant (Caryedes adult) arginyl-tRNA synthetase.
We have used PCR and RACE methods to isolate and characterise the full-length cDNA sequence from Canavalia. The genomic sequence is currently being established. We have compared the derived protein sequence with over 50 sequences in the databases (all non-producers, including Arabidopsis and Rice) and with the crystal structure of the enzyme/arginine complex from yeast. Our results indicate, that the Canavalia sequence is phylogenetically considerably removed from the yeast sequence, so that a modelling on the basis of the yeast structure, while technically possible using the Spdb, need not reflect the true situation in detail. However, the amino acids defined in yeast as being responsible for arginine binding are conserved in Canavalia (as in all other species).
We have been fortunate in establishing a collaboration with the tropical ecologist Dr. D. Janzen (University of Pennsylvania) who is uniquely able to provide specimens of the neotropical bruchid beetle Caryedes brasiliensis and its host plant (Dioclea violacea) from his field studies in Costa Rica, as well as decades of experience with these and other canavanine resistant organisms. He is committed to providing specimens and natural history information from the field, in Costa Rica.
Updated: November 2004 http://www.biologie.uni-freiburg.de/data/bio3/igloi/Accuracy.htm
Perl Molson - 31 Oct 2005 16:19 GMT Later, research of our chair demonstrated that LO exhibited antiviral activity. It inhibited the reproduction of type I Herpes simplex virus. In the presence of 10-2 U/ml (0.3 µg/ml) of the enzyme, the cytopathic effect of type I Herpes simplex virus on the cells cultivated in vitro decreased almost 105-fold. Besides, LO effectively suppresses the expression of the viral antigens. Therefore, it was concluded that highly purified preparations of LO are more effective agents suppressing the reproduction of Herpes simplex virus compared to luteolin and acyclovir [36, 37].
Abbreviations: LO) L-lysine alpha-oxidase.
http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html
Perl Molson - 01 Nov 2005 15:42 GMT Now that's an interesting amino-acid, L-Canavanine (found in alfalfa sprouts, see below). Herpes may take it as L-Arginine. Apparently, Astragalus Root contains L-Canavanine, as well (hmmm, maybe
that's why it works so well against herpes)
http://www.peacefulmind.com/safety.htm ALFALFA (Medicago sativa)
Safety: Alfalfa in its various forms may present some health risks. Powdered alfalfa herb, alfalfa sprouts, and alfalfa seeds all contain L-cavanine, a substance that may cause abnormal blood cell counts, spleen enlargement, or recurrence of lupus in patients with controlled disease. However, heating alfalfa may correct this problem. Researchers investigating alfalfa seeds' ability to lower cholesterol levels discovered that it had another effect on the lab animals used for testing. In some of the monkeys, it caused a disease very similar to lupus. Further research on this effect revealed that monkeys that had abnormal blood cell counts when eating either alfalfa seeds or sprouts, and then recovered when alfalfa was no longer part of their diet, developed the symptoms again when given an isolated component of alfalfa called L-canavanine. Alfalfa seeds and sprouts have a higher concentration of L-canavanine than the leaves or roots. In a clinical trial of alfalfa seeds for lowering cholesterol involving only three human volunteers, one man who participated developed pancytopenia (an abnormally low number of all of the various types of blood cells) and enlargement of the spleen. Additionally, there are two published case reports of patients who had lupus which was controlled with drug therapy, suffering relapses after consuming alfalfa tablets. Again, L-canavanine is thought responsible for these effects. When alfalfa seeds were autoclaved (heated to extremely high temperatures) and fed to monkeys for a year, no ill effects were seen, and the monkeys' cholesterol levels decreased. It may be that the L-canavanine can be destroyed by extreme heat, while the saponins that seem to be responsible for the beneficial effects of alfalfa remain intact. If so a heat-treated product might prove safe; however, much research remains to be done before we can know this for certain. At present, it seems prudent that people who have been diagnosed with lupus, or those who suspect a predisposition to it based on family history, should probably avoid alfalfa. This includes the tablets used for supplements and the sprouts on the salad bar (go for the lettuce or the spinach instead). Because of the estrogenic effects of some of alfalfa's components, alfalfa is not recommended for pregnant or nursing women or young children. In addition, the high vitamin K content in alfalfa could, in theory, make the drug warfarin (Coumadin) less effective. Finally, a number of cases of food poisoning have been documented from fresh sprouts infected with bacteria that was present on the seeds prior to germination. Unfortunately, sprouts can appear fresh and yet host enough bacteria to cause illness in people who eat them. Some health care workers recommend that those at higher risk for such infections-young children, those with chronic diseases, and the elderly-avoid eating sprouts altogether.
Dosage:A typical dose of alfalfa for tea is 1 to 2 teaspoons per cup, steeped in boiling water for 10 to 20 minutes. Tablets and capsules of whole alfalfa or alfalfa extracts should be taken according to the manufacturer's recommendations. Certain products are said to be free of canavanine and other potentially harmful constituents, and may be preferable.
Contraindications: If you are taking warfarin (Coumadin), the high vitamin K content of alfalfa might make it less effective.
NATURAL TOXINS IN BEANS ARE NO CAUSE FOR ALARM by Steve Meyerowitz People have been alarmed by articles alleging that alfalfa and other sprouts contain toxins. This news has struck fear in Vegetarians and others because sprouts have always been considered "health food." Most of us have heard this news from the very popular author and proponent of natural healing, Andrew Weil. When questioned for more detail, Dr. Weil refers us to well known toxicologist Bruce Ames. Ames has written extensively on plant toxins. In a recent telephone interview, Dr. Ames claimed that he is not against sprouts. "Eat any sprouts you want..." he said. "There's nothing wrong with sprouts....If I was to be against sprouts then I would be against every kind of vegetation."
Ames tells us that there are toxins in the seeds of many common plants and vegetables as a natural protective mechanism against insects and other predators. The natural toxin identified in sprouting seeds is known as L-canavanine, a pre-cursor necessary for the development of the amino acid arginine. L-canavanine is one of 600 non-protein amino acids synthesized by plants. It is found in hundreds of legumes (beans) and other arginine rich foods such as garlic and onion.
However, it's presence in beans (alfalfa is a tiny bean) is not a cause for alarm. During the germination process, L-Canavanine is converted to Arginine, one of the essential amino acids. In 1960, Dr. E.A. Bell, demonstrated that once alfalfa reaches the green stage, this toxin is reduced to a trace amount. More recently, L-Canavanine biochemist Gerald A. Rosenthal of the University of Kentucky found that this natural toxin "was shown to exhibit significant antineoplastic activity against MIAPaCa-2, a human pancreatic cancer." This natural plant protectant can also protect us against cancer! This work is underscored by that of John Hopkins researcher Paul Talalay who in 1997 found that the cancer protectant sulphoraphane is present in broccoli sprouts in quantities up to 50 times greater than in broccoli.
The research quoted on this subject involved feeding monkeys L-canavanine sulfate tablets plus biscuits made from raw, unsprouted alfalfa beans and "sprouts." However, none of this research actually involved alfalfa sprouts as they are typically consumed by people. The "sprouts" were beans germinated for 1-3 days and then oven dried. They never turned green. Sprouts for human consumption are grown for approximately 7 days and contain only a trace of L-canavanine.
The thrust of the research was to explore the connection between this toxin and the auto-immune disease Lupus. It was not a test of alfalfa sprouts. In fact, "sprouts" played only a marginal role in 2 of the numerous studies on this toxin between done between 1981 and 1984. According to Lupus specialist Dr. Emil J. Bardana, who worked with Dr. Rene Malinow on the two studies involving the so called "sprouts," there is no basis to say that eating alfalfa sprouts would cause lupus or SLE. "I wouldn't discourage my lupus patients from eating alfalfa sprouts." The amount of sprouts you eat on a salad "isn't anywhere near the dose we fed the monkeys."
Keeping things in perspective, many common foods contain toxins all of which are consumed in greater quantities than sprouts: potatoes have solanine, mushrooms have strychnine, apples and apricots have cyanide. If you believe such toxins threaten our health, then you should also eliminate tomatoes, eggplants and peppers, mustard, cottonseed oil, peanuts, black pepper, root beer (sassafras) and last but not least, coffee which, according to Dr. Ames, contains the "natural mutagen chlorogenic acid and highly toxic atractylosides".
Let us not forget that as living plants, sprouts are some of our richest sources of phytochemicals, enzymes, anti-oxidants, nitrosamines, trace minerals and chemo-protectants such as sulphoraphane and isoflavone which work against toxins, resist cell mutation and invigorate the body's immune system. Don't doubt the sprout!
"Until Man duplicates a blade of grass, nature can laugh at his so called scientific knowledge." --Thomas Edison
©1998 Steve Meyerowitz
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http://www.sproutman.com/toxins.html
> Later, research of our chair demonstrated that LO exhibited antiviral > activity. It inhibited the reproduction of type I Herpes simplex virus. [quoted text clipped - 9 lines] > > http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html Perl Molson - 02 Nov 2005 17:03 GMT Some additional readings:
http://www.pubmedcentral.nih.gov/pagerender.fcgi?artid=1066589&pageindex=1#page
l-Arginine and l-Canavanine Metabolism in Jack Bean, Canavalia ensiformis (L.) DC. and Soybean, Glycine max (L.) Merr. 1
http://www.protein.bio.msu.ru/biokhimiya/contents/v67/pdf/bcm_1152.pdf LLysine aOxidase: Physicochemical and Biological Properties
> Now that's an interesting amino-acid, L-Canavanine (found in alfalfa > sprouts, see below). [quoted text clipped - 152 lines] > > > > http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html
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