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Phytohaemagglutinin (PHA, or phytohemagglutinin) is a
lectin Lectins are carbohydrate-binding proteins that are highly specific for sugar groups that are part of other molecules, so cause agglutination of particular cells or precipitation of glycoconjugates and polysaccharides. Lectins have a role in rec ...
found in plants, especially certain
legumes A legume () is a plant in the family Fabaceae (or Leguminosae), or the fruit or seed of such a plant. When used as a dry grain, the seed is also called a pulse. Legumes are grown agriculturally, primarily for human consumption, for livestock for ...
. PHA actually consists of two closely related proteins, called leucoagglutinin (PHA-L) and PHA-E. These proteins cause blood cells to clump together. PHA-E cause
erythrocytes Red blood cells (RBCs), also referred to as red cells, red blood corpuscles (in humans or other animals not having nucleus in red blood cells), haematids, erythroid cells or erythrocytes (from Greek ''erythros'' for "red" and ''kytos'' for "holl ...
(red blood cells) to clump. PHA-L causes
leukocytes White blood cells, also called leukocytes or leucocytes, are the cells of the immune system that are involved in protecting the body against both infectious disease and foreign invaders. All white blood cells are produced and derived from mu ...
(white blood cells) to clump. Phytohaemagglutinin has carbohydrate-binding specificity for a complex oligosaccharide containing
galactose Galactose (, '' galacto-'' + ''-ose'', "milk sugar"), sometimes abbreviated Gal, is a monosaccharide sugar that is about as sweet as glucose, and about 65% as sweet as sucrose. It is an aldohexose and a C-4 epimer of glucose. A galactose molecu ...
, ''N''-acetylglucosamine, and
mannose Mannose is a sugar monomer of the aldohexose series of carbohydrates. It is a C-2 epimer of glucose. Mannose is important in human metabolism, especially in the glycosylation of certain proteins. Several congenital disorders of glycosylat ...
. It is found in the highest concentrations in uncooked red
kidney bean The kidney bean is a variety of the common bean ('' Phaseolus vulgaris''). It resembles a human kidney and thus is named after such. Red kidney beans should not be confused with other red beans, such as adzuki beans. Classification There ar ...
s and white kidney beans (also known as
cannellini The kidney bean is a variety of the common bean (''Phaseolus vulgaris''). It resembles a human kidney and thus is named after such. Red kidney beans should not be confused with other red beans, such as adzuki beans. Classification There are ...
), and it is also found in lower quantities in many other types of
green bean Green beans are young, unripe fruits of various cultivars of the common bean ('' Phaseolus vulgaris''), although immature or young pods of the runner bean ('' Phaseolus coccineus''), yardlong bean ( ''Vigna unguiculata'' subsp. ''sesquipedali ...
s and other common beans (''
Phaseolus vulgaris ''Phaseolus vulgaris'', the common bean, is a herbaceous annual plant grown worldwide for its edible dry seeds or green, unripe pods. Its leaf is also occasionally used as a vegetable and the straw as fodder. Its botanical classification, ...
''), as well as broad beans (''
Vicia faba ''Vicia faba'', commonly known as the broad bean, fava bean, or faba bean, is a species of vetch, a flowering plant in the pea and bean family Fabaceae. It is widely cultivated as a crop for human consumption, and also as a cover crop. Varieti ...
'') such as fava beans. It has a number of physiological effects and is used in medical research. In high doses, it is a
toxin A toxin is a naturally occurring organic poison produced by metabolic activities of living cells or organisms. Toxins occur especially as a protein or conjugated protein. The term toxin was first used by organic chemist Ludwig Brieger (1849 ...
. The lectin has a number of effects on cell metabolism; it induces
mitosis In cell biology, mitosis () is a part of the cell cycle in which replicated chromosomes are separated into two new nuclei. Cell division by mitosis gives rise to genetically identical cells in which the total number of chromosomes is maintai ...
, and affects the cell membrane in regard to transport and permeability to proteins. It agglutinates most mammalian
red blood cell Red blood cells (RBCs), also referred to as red cells, red blood corpuscles (in humans or other animals not having nucleus in red blood cells), haematids, erythroid cells or erythrocytes (from Greek ''erythros'' for "red" and ''kytos'' for "hol ...
types. As a toxin, it can cause poisoning in
monogastric A monogastric organism has a simple single-chambered stomach (one stomach). Examples of monogastric herbivores are horses and rabbits. Examples of monogastric omnivores include humans, pigs, hamsters and rats. Furthermore, there are monogastric ...
animals, such as humans, through the consumption of raw or improperly prepared legumes, ''e.g.'', beans. Measured in haemagglutinating units (hau), a raw red kidney bean may contain up to 70,000 hau, but this is reduced to between 200 and 400 hau when properly cooked. Studies by British scientists recommend soaking beans for at least five hours, discarding the water, and then boiling the beans in fresh water at for at least thirty minutes. A pressure cooker at 15 psi may be used to cook beans in 45 minutes without presoaking. Insufficient cooking, such as in a
slow cooker A slow cooker, also known as a crock-pot (after a trademark owned by Sunbeam Products but sometimes used generically in the English-speaking world), is a countertop electrical cooking appliance used to simmer at a lower temperature than o ...
at 75 °C/ 167 °F, may not completely destroy the toxins. Beans also contain
alpha amylase inhibitor In molecular biology, alpha-amylase inhibitor (or α-...) is a protein family which inhibits mammalian alpha-amylases specifically, by forming a tight stoichiometric 1:1 complex with alpha-amylase. This family of inhibitors has no action on plant ...
, but not in sufficient quantities to affect the digestion of starch after consumption of beans. Poisoning can be induced from as few as four raw beans. Symptoms usually begin with extreme nausea and vomiting within one to three hours of ingestion, followed by diarrhea. Abdominal pain has been reported in some people. Recovery is usually spontaneous and rapid, occurring within three to four hours after onset of symptoms, although some cases have required hospitalization. In medicine these proteins are useful and are used as a
mitogen A mitogen is a small bioactive protein or peptide that induces a cell to begin cell division, or enhances the rate of division (mitosis). Mitogenesis is the induction (triggering) of mitosis, typically via a mitogen. The mechanism of action of a ...
to trigger T-lymphocyte cell division and to activate latent HIV-1 from human peripheral
lymphocyte A lymphocyte is a type of white blood cell (leukocyte) in the immune system of most vertebrates. Lymphocytes include natural killer cells (which function in cell-mediated, cytotoxic innate immunity), T cells (for cell-mediated, cytotoxic a ...
s. In
neuroscience Neuroscience is the science, scientific study of the nervous system (the brain, spinal cord, and peripheral nervous system), its functions and disorders. It is a Multidisciplinary approach, multidisciplinary science that combines physiology, an ...
,
anterograde tracing In neuroscience, anterograde tracing is a research method which is used to trace axonal projections from their source (the cell body or soma) to their point of termination (the synapse). A hallmark of anterograde tracing is the labeling of the presy ...
is a research method that uses the protein product phytohaemagglutinin PHA-L as a molecular tracer that can be taken up by the cell and transported across the synapse into the next cell thereby tracing the path of axonal projections and relative connections that nerve impulses travel beginning with the source located at the
perikaryon The soma (pl. ''somata'' or ''somas''), perikaryon (pl. ''perikarya''), neurocyton, or cell body is the bulbous, non-process portion of a neuron or other brain cell type, containing the cell nucleus. The word 'soma' comes from the Greek '' σῶ ...
(cell body or soma) and through the presynaptic part located on
neuron A neuron, neurone, or nerve cell is an electrically excitable cell that communicates with other cells via specialized connections called synapses. The neuron is the main component of nervous tissue in all animals except sponges and placozoa ...
's efferent
axon An axon (from Greek ἄξων ''áxōn'', axis), or nerve fiber (or nerve fibre: see spelling differences), is a long, slender projection of a nerve cell, or neuron, in vertebrates, that typically conducts electrical impulses known as action p ...
all the way to the point of termination at the efferent synapse which then provides input to another neuron. Lymphocytes cultured with phytohaemagglutinin can be used for
karyotype A karyotype is the general appearance of the complete set of metaphase chromosomes in the cells of a species or in an individual organism, mainly including their sizes, numbers, and shapes. Karyotyping is the process by which a karyotype is disce ...
analysis. Stimulation of peripheral blood lymphocytes by phytohaemagglutinin presents a classic model of transition of cells from the quiescent G0 phase of the cell cycle into G1-, and subsequently progression through S-, G2- and M- phases of the cycle.


History

Prior to 1960, crude extracts of PHA were known to coat the surface of red blood cells, make them heavier, and thereby improve the separation of the white cell buffy coat. Peter Nowell, an immunologist and pathologist at the University of Pennsylvania in Philadelphia, was using PHA for this purpose in 1960 when he discovered it also had the ability to stimulate mitotic division of lymphocytes from normal peripheral blood. Prior to his discovery, these cells were assumed to be the terminal end-products of differentiation. This work had tremendous implications for the constitutional study of chromosomal disorders. T. C. Hsu's 1979 book "Human and Mammalian Cytogenetics an Historical Perspective" (ISBN 978-1-4612-6159-9) is an excellent resource for this history.


References


External links

* {{Lectins Proteins Lectins Foodborne illnesses Legume lectins