ALBERT

Notes: Galactosephilic and mannosephilic lectins from Pseudomonas aeruginosa interact with Tetrahymena pyriformis GL. Specific adsorption of these lectins onto the Tetrahymena can be shown by inhibition of hemagglutination and by peroxidase binding to the cells mediated by the mannosephilic lectins. Interaction with the lectins does not agglutinate the protozoa even after immobilization by Na fluoride, formaldehyde, and glutaraldehyde or after papain treatment. However, inclusion of the lectins in the growth medium increases the growth rate of Tetrahymena and their presence in the medium supplied to starved ciliates increases phagocytosis of Chinese ink and vacuolization.

Notes: SYNOPSIS The galactosephilic and mannosephilic hemagglutinins of Pseudomonas aeruginosa adsorbed onto Euglena gracilis, Chlamydomonas reinhardi, and Tetrahymena pyriformis. Furthermore, peroxidase binding to the 3 protozoan species was shown to be mediated by these lectins. Binding of Pseudomonas lectins to E. gracilis and C. reinhardi caused their specific agglutination, whereas no agglutination was observed with T. pyriformis, even after treatment by papain or by NaF. Added to the culture medium, the Pseudomonas hemagglutinins stimulated growth of E. gracilis and T. pyriformis due to their binding to these protozoa: this effect was partly inhibited by the specific sugar.

Notes: The two Pseudomonas aeruginosa lectins PA-IL and PA-IIL, which are very similar in subunit size, composition and properties, but differ in carbohydrate specificity, were shown to exhibit opposite temperature profiles in hemagglutination tests. The galactophilic PA-IL, which interacts with the erythrocyte I antigen (together with B or P system antigens), resembles Ii system-specific ‘cold hemagglutinins’ (including antibodies and lectins of animals and plants) in low (4°C) temperature optimum, while the hemagglutination by the fucose- and mannose-binding PA-IIL (like that of antibodies and lectins which do not bind to these antigens) increases on raising the temperature from 4 to 37°C and even to 42°C. The preferential production of both P. aeruginosa lectins at 28°C and their much stronger interaction with enzyme (protease or sialidase)-damaged cells, as well as the lower temperature optimum (4°C) of PA-IL-binding to the host cells, may be associated with the saprophytic rather than parasitic designation of this bacterium.

Notes: Using the 33 N-terminal amino acids of the fucose/mannose binding lectin PA-IIL of Pseudomonas aeruginosa ATCC 33347 in a tblastn search of P. aeruginosa PAO1 genomic sequence in GenBank revealed a single open reading frame encoding a 114-amino acid protein (excluding initiator methionine) perfectly matching that amino acid sequence. Following its stop codon there is a GC-rich sequence having a perfect dyad symmetry promoting formation of a hairpin loop structure, potentially enabling rho-independent transcription termination. Upstream of the putative ribosomal binding site there are sequences resembling Vibrio fischeri luxI box, consistent with autoinduction of this gene. The predicted PA-IIL molecular mass, confirmed by mass spectrometry, is 11 732 Da. Its pI is 3.88. The C-terminal domain is particularly hydrophobic, implying possible embedding in the cell membrane. PA-IIL is similar to P. aeruginosa PA-IL lectin in some amino acids and potential glycosylation sites but lacks cysteine, methionine and histidine. Despite their relations in functions and regulation, their genes are widely separated (by about 867.5 kb).

Notes: Lectins are important tools for cell typing and for the study of cell surface components. They have been widely used for the analysis of carbohydrates on the surface of many eukaryotic and prokaryotic cells, but they have not yet been exploited in the study of the halophilic Archaea (family Halobacteriaceae), because of the high salinity required for the structural integrity of these microorganisms. We have defined the salt concentration threshold high enough for survival of the Archaea, but sufficiently low for lectins to bind to them. Under these conditions we studied the interactions of a series of lectins, exhibiting different sugar specificities, with diverse halophilic Archaea. Concanavalin A was the most reactive by virtue of its glucose (and mannose) binding. The other lectins varied in their interactions. The results indicate that lectins might be useful probes for both archaeal typing and analysis of their cell surface carbohydrates.

Notes: Aplysia gonad lectin (AGL), which strongly agglutinates cancer cells, was found, in the present study, to bind to erythrocyte T antigen, in addition to its affinity to Ii system antigens. These antigens were reported to be overexpressed and to contribute to tumor progression and invasion. In healthy human sera, there are antibodies against them, stimulated by the normal intestinal microflora, which bear similar glycoforms. Since the levels of these antibodies were reported to be lower in most cancer patients’ sera, we have examined the applicability of AGL to isolation of enteric commensal Escherichia coli strains which bear glycoforms cross-reacting with the cancer-associated antigens. Among 30 E. coli isolates examined, two were agglutinated by AGL. One of them was also agglutinated by certain related galactophilic lectins, which bind to the T and Tn antigens. The agglutination of the two bacteria by healthy human sera, as a group, was stronger than that displayed by the cancer patients’ sera. These results indicate that AGL might be useful for identification of the desired bacteria, which could potentially serve for cancer diagnosis and therapy.

Notes: Pseudomonas aeruginosa produces a galactophilic lectin, PA-IL, that resembles P-fimbrial adhesins of uropathogenic Escherichia coli strains in binding to human P blood group antigens. We examined, in the present study, its interaction with pigeon egg white glycoproteins carrying N-glycans with terminal Galα1–4Gal which inhibit the adhesion of P-fimbriae. For comparison, the lectin concanavalin A (Con A) and additional avian egg whites (of hen and quail) were also examined. The results obtained in both hemagglutination inhibition and Western blot analyses showed that PA-IL, unlike Con A, preferentially reacted with the pigeon egg white glycoproteins. These results, which confirmed PA-IL similarity in sugar specificity to E. coli P-fimbriae, demonstrated the advantage of this purified lectin for representing P-type and additional galactophilic microbial adhesins unavailable in purified stable form, in Western blot analyses.

Notes: Abstract Lectins are ubiquitous proteins, which exhibit a specific and reversible sugar-binding activity. They react with glycosylated macromolecules and cells and may coaggregate them and lead to their lysis or alterations. Various lectin biological effects are well known, but their basic biological function is considered as yet unknown. In the present review, an experimental evidence and theoretical considerations are forwarded for supporting our suggestion that the general basic lectin or lectinoid (lectin-like protein) function in microorganisms, plants and animals is a cofunction enabling the activities of key lytic enzymes (lysins: glycosidases, proteases, esterases, phosphatases, hemolysin, etc.). The lectin service is: homing onto glycosylated receptors, anchoring to them and induction of cooperative conformational effects which enable their counterpart lysin activity on exogenous or endogenous target molecules and cells. The ‘lectin-lysin’ pair may reside in the same molecule, or in linked subunits. It may also be formed by cofunction of two separate entities originating from one or two (homogenous or heterogenous) cell sources. The lectin and lysin may be free or cell-bound components located intra or extracellularly. The final result of their cofunction is practically irreversible; either cell and macromolecule lysis for nutrition, homeostasis and protection or cell alteration, reorganization and new productivity.Our suggestion emphasizes the prominent analogy of lectins to lytic enzyme positioning sites (LEPS), immunoglobulins and polypeptide hormones. The lectin analogy to LEPS and immunoglobulins is exhibited in the lectin-dependent cell and macromolecule lysis for nutritional and homeostatic purposes or for protection, respectively. The hormone-like lectin activity is exhibited in the lectin-dependent cell alterations. In addition to similar functions and effects, the analogy also includes the properties and behavior of these proteins. The suggested hypothesis is based on experimental evidence from microorganisms, plants and animals. It envisions the lectin and lectinoid function in cell attacks on glycosylated molecules or cells, cell-substratum and cell-cell interactions (fusion, invasion, etc.), cell transformation and formation of special structures. All of them according to a developmental program, or special (especially unfavourable) environmental conditions. The lectin resistance to proteolysis and unfavourable pH or temperature is in accord with the suggested hypothesis.

Notes: The ability of Pseudomonas aeruginosa purified lectin vaccine to protect ICR mice against lethal infection caused by various serotypes of this bacterium was examined. Intraperitoneal injection of the lectin preparation was effective in full protection of the mice against the homologous strain. It was also effective in conferring from a partial to a full protection against most of the other virulent strains tested. The only strain which was still lethal for the immunized mice was one which produces exotoxin A but not lectins.