Structural Components Of Cell Wall Research Articles

DNA aptamers bind specifically and selectively to (1 → 3)-β- d-glucans

(1 → 3)-β- d-Glucans are structural cell wall components of fungi, plants, and some bacteria and have been linked with human respiratory symptoms following aerosol exposure. A clear interpretation of the health impact of (1 → 3)-β- d-glucans is limited by the high cost and uncertainties associated with current glucan quantitation methods. The objective of this research is to develop DNA aptamers for the measurement of (1 → 3)-β- d-glucans. Aptamers are synthetic DNA functional binding molecules that fold into unique conformations, allowing them to bind specifically to their target. Through the in vitro selection process SELEX, we have produced aptamers that are able to bind with sub-micromolar affinity to curdlan, a linear unbranched form of (1 → 3)-β- d-glucans. These aptamers display high selectivity to curdlan and do not bind to non-(1 → 3)-β- d-polysaccharides, suggesting specificity for the β-(1 → 3)-glycosidic linkage. The aptamers produced here will enable the production of more cost-effective, less ambiguous assays for the environmental measurement of (1 → 3)-β- d-glucans.

Contribution of dectin-1 and granulocyte macrophage–colony stimulating factor (GM-CSF) to immunomodulating actions of β-glucan

β-Glucans are major cell wall structural components in fungi. As they are not found in animals, these carbohydrates are considered to be classic pathogen-associated molecular patterns (PAMPs), and are recognized by the innate immune system. Although their immunomodulating activities have been shown to be associated with the recognition of some fungi, and with their medicinal properties in the field of cancer immunotherapy, it is still unclear how β-glucans mediate their effects. Recent studies have started to shed some light on their cellular receptors, such as dectin-1, and their molecular mechanisms of action. We have extensively investigated the response of leukocytes to β-glucan, focusing on cytokine induction by SCG, which is a major 6-branched 1,3-β-d-glucan in Sparassis crispa Fr. There is a strain difference in the reactivity of mice to SCG, and DBA/1 and DBA/2 mice are highly sensitive strains. In the process of research on cytokine induction by SCG in DBA/2 mice, we found that GM-CSF plays a key biological role in this activity. Cytokine induction by SCG was completely abolished in dendritic cells from dectin-1 knockout mice. On the other hand, controlling the level of endogenous GM-CSF production and/or dectin-1 expression could regulate the reactivity to β-glucan. These results indicate that the key factors in the responsiveness to β-glucan are GM-CSF production and dectin-1 expression. In this review, we describe how the key molecules related to the expression of the immunomodulating activities of β-glucan were identified, and how the response to β-glucan is controlled.

The potential of Raman microscopy and Raman imaging in plant research

To gain a better understanding on structure, chemical composition and properties of plant cells, tissues and organs several microscopic, chemical and physical methods have been applied during the last years. However, a knowledge gap exists about the location, quantity and structural arrangement of molecules in the native sample or what happens on the molecular level when samples are chemically or mechanically treated or how they respond to mechanical stress. These questions need to be answered to optimise utilization of plants in food industry and pharmacy and to understand structure-function relationships of plant cells to learn from natures unique. Advances in combining microscopy with Raman spectroscopy have tackled this problem in a non-invasive way and provide chemical and structural informationin situwithout any staining or complicated sample preparation. In this review the different Raman techniques (e.g. near infrared Fourier Transform Raman spectroscopy (NIR-FT), resonance Raman spectroscopy, surface-enhanced Raman spectroscopy) are briefly described before approaches in plant science are summarised. Investigations on structural cell wall components, valuable plant substances, metabolites and inorganic substances are included with emphasis on Raman imaging. The introduction of the NIR-FT-Raman technique led to many applications on green plant material by eliminating the problem of sample fluorescence. For mapping and imaging of whole plant organs (seeds, fruits, leaves) the lateral resolution (~10μm) of the NIR-FT technique is adequate, whereas for investigations on the lower hierarchical level of cells and cell walls the high resolution gained with a visible laser based system is needed. Examples on high resolution Raman imaging are given on wood cells, showing that changes in chemistry and orientation can be followed within and between different cell wall layers having dimensions smaller than 1 μm. In addition imaging the distribution of amorphous silica is shown on horsetail tissue, including an area scan from a cross section as well as a depth profiling within a silica rich knob of the outer stem wall.

Purification and characterization of proline/hydroxyproline-rich glycoprotein from pearl millet coleoptiles infected with downy mildew pathogen Sclerospora graminicola

Hydroxyproline-rich glycoproteins (HRGPs) are important plant cell wall structural components, which are also involved in response to pathogen attack. In pearl millet, deposition and cross-linking of HRGPs in plant cell walls was shown to contribute to the formation of resistance barriers against the phytopathogenic oomycete Sclerospora graminicola. In the present study, the purification and characterization of HRGPs that accumulated in coleoptiles of pearl millet seedlings in response to S. graminicola inoculation has been carried out. Periodic acid Schiff’s staining revealed that the purified protein was a glycoprotein. The protein to carbohydrate ratio was determined to be 95.5%:4.5% (w/w). Proline amounted for 20 mol% of the total amino acids as indicated by amino acid composition analysis. The isolated protein had a p I of 9.8 and was shown to be composed of subunits of 27, 17, and 14 kDa. Cross reactivity with the monoclonal antibody MAC 265 and the presence of the signature amino acid sequence, PVYK, strongly suggested to classify the purified glycoprotein as a member of the P/HRGPs class. In the presence of horseradish peroxidase and H 2O 2 the purified glycoprotein served as a substrate for oxidative cross-linking processes.

Diferulate Content of Maize Sheaths Is Associated with Resistance to the Mediterranean Corn Borer Sesamia nonagrioides (Lepidoptera: Noctuidae)

The leaf sheaths of selected inbred lines of maize (Zea mays L.) with variable levels of stem resistance to the Mediterranean corn borer Sesamia nonagrioides (Lefèvbre) were evaluated for antibiotic effect on insect development. Phytochemical analyses of leaf sheaths were conducted for cell wall phenylpropanoid content to gain a better understanding of maize-resistance mechanisms. Laboratory bioassays established that sheath tissues from different genotypes significantly affected the growth of neonate larvae. Three hydroxycinnamates, p-coumaric, trans-ferulic, and cis-ferulic acids, and three isomers of diferulic acid, 8-5', 8-O-4', and 8-5' b (benzofuran form), were identified. Significant negative correlations were found between larvae weight and diferulic acid content for six genotypes. These results are in agreement with previous studies concerning the role of cell wall structural components in stem borer resistance.

Orally administered marine (1 → 3)-β- d-glucan Phycarine stimulates both humoral and cellular immunity

(1 → 3)-β- d-Glucans represent highly conserved structural components of cell walls in yeast, fungi, or seaweed. However, it is still unknown how they mediate their effects. The aim of this study was to evaluate both intraperitoneal and oral application of seaweed-derived (1 → 3)-β- d-glucan Phycarine. Phycarine showed significant stimulation of phagocytosis by peripheral blood cells. In addition, the efficiency of chemotherapy of Lewis lung carcinoma with cyclophosphamide was potentiated by Phycarine administration. Phycarine also strongly shortened the recovery of leucopenia caused either by chemotherapy or irradiation. Besides the role in stimulation of cellular immunity, we also found a significant increase of antibody formation. Using a suckling rat model for evaluation of the absorption and tissues distribution of enterally administered 125I-Phycarine, we found that the majority of Phycarine was detected in the stomach and duodenum 5 min after the administration. This amount sharply decreased during first 30 min. A significant amount of Phycarine entered proximal intestine in a shortly after the gavage. Its transit through proximal intestine was decreasing with time and simultaneously increasing in the ileum. Systemic blood levels were very low (less than 0.5%). Taken together, these observations suggest that Phycarine is similarly effective both after i.p. and oral application, has very strong stimulating effects on three types of experimentally induced leucopenia and stimulates both humoral and cellular branch of immune reactions. The majority of Phycarine can be detected throughout the gastrointestinal tract, supporting the feasibility of enteral administration of Phycarine in the treatment of gastrointestinal diseases.

(13)-beta-D-glucans and respiratory health: a review of the scientific evidence

(1-->3)-Beta-D-glucan are non-allergenic structural cell wall components of most fungi that have been suggested to play a causal role in the development of respiratory symptoms associated with indoor fungal exposure. This review describes the currently available epidemiological literature on health effects of (1-->3)-beta-D-glucan, focusing on atopy, airway inflammation and symptoms, asthma, and lung function. In addition to population studies, studies in human volunteers experimentally exposed to (1-->3)-beta-D-glucan are described as well as relevant animal studies. Furthermore, the review discusses exposure assessment methods, the potential for exposure control and it concludes with identifying research needs. The observational and experimental studies reviewed suggested some association between (1-->3)-beta-D-glucan exposure, airway inflammation and symptoms, however, results were mixed and specific symptoms and potential underlying inflammatory mechanisms associated with exposure could not be identified. Large observational studies using well validated exposure assessment methods are needed to further our knowledge regarding the potential health effects of indoor (1-->3)-beta-D-glucan exposure. The currently available epidemiological data do not permit conclusions to be drawn regarding the presence (or absence) of an association between environmental (1-->3)-beta-D-glucan exposure and specific adverse health effects, nor is it clear from the currently available evidence which specific immunological mechanisms underlie the presumed health effects. More and larger observational studies are needed to asses whether (1-->3)-beta-D-glucan exposure plays a significant role in respiratory morbidity. In addition, existing methods to assess environmental (1-->3)-beta-D-glucan exposure require validation and further development before they can be used routinely in large scale epidemiological studies.

Coccidioides posadasii Contains a Single 1,3-β-Glucan Synthase Gene That Appears To Be Essential for Growth

1,3-beta-Glucan synthase is responsible for the synthesis of beta-glucan, an essential cell wall structural component in most fungi. We sought to determine whether Coccidioides posadasii possesses genes homologous to known fungal FKS genes that encode the catalytic subunit of 1,3-beta-glucan synthase. A single gene, designated FKS1, was identified, and examination of its predicted protein product showed a high degree of conservation with Fks proteins from other filamentous fungi. FKS1 is expressed at similar levels in mycelia and early spherulating cultures, and expression decreases as the spherules mature. We used Agrobacterium-mediated transformation to create strains that harbor DeltaFKS1::hygB, a null allele of FKS1, and hypothesize that Fks1p function is essential, due to our inability to purify this allele away from a complementing wild-type FKS1 allele in a heterokaryotic strain. The heterokaryon appears normal with respect to growth rate and arthroconidium production; however, microscopic examination of strains with DeltaFKS1::hygB alleles revealed abnormal swelling of hyphal elements.

Accumulation of hydroxyproline-rich glycoproteins in pearl millet seedlings in response to Sclerospora graminicola infection

Hydroxyproline-rich glycoproteins (HRGPs) are important plant cell wall structural components that are known to be induced in response to pathogen attack. In the present study, accumulation of HRGPs in pearl millet seedlings in response to Sclerospora graminicola, the incitant of downy mildew disease was monitored by estimation of hydroxyproline (Hyp) in the hydrolyzed cell wall samples. In 2-day-old seedlings of a highly resistant cultivar of pearl millet inoculated with S. graminicola, a marked increase in Hyp content was observed in the coleoptiles as early as 2 h compared to that in a highly susceptible cultivar. A correlation was observed between the amount of Hyp induced in coleoptiles of different cultivars of pearl millet to S. graminicola infection and the host resistance. Electrophoretic separation of the acid–ethanol extracted proteins from these coleoptiles followed by periodic acid Schiff staining revealed three glycoproteins of molecular masses 27, 17 and 14 kDa. These three glycoproteins also reacted positively on western blots with MAC 265, a monoclonal antibody against pea HRGP, indicating that they are HRGPs. The 17 kDa HRGP was markedly increased in highly resistant cultivar to S. graminicola infection compared to highly susceptible cultivar. The 14 kDa HRGP was exclusively present in the highly resistant cultivar and not in highly susceptible cultivar. Histological studies showed cross-linking of proteins in the cell walls of only highly resistant cultivar against S. graminicola infection. In addition, immunocytochemical studies using MAC 265 antibodies recognized these cross-linked proteins to be HRGPs. These findings indicated that accumulation of HRGPs in cell walls of highly resistant pearl millet seedlings is a component of defense reaction against downy mildew pathogen S. graminicola.

Sequential fractionation and two-dimensional gel analysis unravels the complexity of the dimorphic fungus Candida albicans cell wall proteome.

The cell wall proteins of Candida albicans play a key role in morphogenesis and pathogenesis and might be potential target sites for new specific antifungal drugs. However, these proteins are difficult to analyze because of their high heterogeneity, interconnections with wall polysaccharides (mannan, glucan, and chitin), low abundance, low solubility, and hydrophobic nature. Here we report a subproteomic approach for the study of the cell wall proteins (CWPs) from C. albicans yeast and hyphal forms. Most of the mannoproteins present in this compartment were extracted by cell wall fractionation according to the type of interactions that they establish with other structural components. CWPs were solubilized from isolated cell walls by hot SDS and dithiothreitol treatment followed by extraction either by mild alkali conditions or by enzymatic treatment with glucanases and chitinases. These highly enriched cell wall fractions were analyzed by two-dimensional PAGE, showing that a large number of proteins are involved in cell wall construction and that the wall remodeling that occurs during germ tube formation is related to changes in the composition of CWPs. We suggest that the CWP-chitin linkage is an important retention mechanism of CWPs in C. albicans mycelial forms. This article also highlights the usefulness of the combination of sequential fractionation and two-dimensional PAGE followed by Western blotting using specific antibodies against known CWPs in the characterization of incorporation mechanisms of such CWPs into the cell wall and of their interactions with other wall components. Mass spectrometry analyses have allowed the identification of several cell surface proteins classically associated with both the cell wall and other compartments. The physiological significance of the dual location of these moonlighting proteins is also discussed. This approach is therefore a powerful tool for obtaining a comprehensive and integrated view of the cell wall proteome.

Isolation and characterization of holocellulose from Alfalfa

Holocellulose isolated from the aerial parts of alfalfa (Medicago sativa) contains a polysaccharide complex of cellulose and hemicelluloses, the major structural components of cell walls. Holocellulose is highly hydrophilic and has a dense biopolymer packing. The carboxylic groups of hemicelluloses and cellulose determines the ability of holocellulose to adsorb polyvalent metal cations.

A dynamical model for plant cell wall architecture formation.

We discuss a dynamical mathematical model to explain cell wall architecture in plant cells. The highly regular textures observed in cell walls reflect the spatial organisation of the cellulose microfibrils (CMFs), the most important structural component of cell walls. Based on a geometrical theory proposed earlier [A. M. C. Emons, Plant, Cell and Environment 17, 3-14 (1994)], the present model describes the space-time evolution of the density of the so-called rosettes, the CMF synthesizing complexes. The motion of these rosettes in the plasma membrane is assumed to be governed by an optimal packing constraint on the CMFs plus adherent matrix material, that couples the direction of motion, and hence the orientation of the CMF being deposited, to the local density of rosettes. The rosettes are created inside the cell in the endoplasmatic reticulum and reach the cell-membrane via vesicles derived from Golgi-bodies. After being inserted into the plasma membrane they are assumed to be operative for a fixed, finite lifetime. The plasma membrane domains within which rosettes are activated are themselves also supposed to be mobile. We propose a feedback mechanism that precludes the density of rosettes to rise beyond a maximum dictated by the geometry of the cell. The above ingredients lead to a quasi-linear first order PDE for the rosette-density. Using the method of characteristics this equation can be cast into a set of first order ODEs, one of which is retarded. We discuss the analytic solutions of the model that give rise to helicoidal, crossed polylamellate, helical, axial and random textures, since all cell walls are composed of (or combinations of) these textures.

Soluble Polysaccharide Content at Initial Time of Experimental Must Preparation

The diversity of procedures used for the analysis of polysaccharides in musts obtained at laboratory scale did not allow their accurate quantification and characterization. Soluble polysaccharides have been isolated from musts at the initial time of preparation of Sauvignon blanc and Ugni blanc cultivars and the influence of the mode of preparation has been investigated. Heat treatment, commonly used for the inactivation of endogenous glycosylhydrolases, led to an overestimation of polysaccharide contents. Consequently, the characterization of must polysaccharides requires their isolation from crushed and pressed grapes, a procedure representative of the typical process used for white winemaking. Glycosyl residue, determined by GC of their per-<i>O</i>trimethylsilylated methyl glycosides, and glycosyl linkage, determined by GC of their partially methylated and carboxyl-reduced alditol acetates, composition analyses showed that polysaccharides were a complex mixture of pectic (galacturonans, type I and II arabinogalactans, arabinans), cellulosic and hemicellulosic (xyloglucans and mannans) polysaccharides. Type II arabinogalactan proteins, which are soluble proteoglycans, are the most abundant polysaccharides, whereas the others, that are structural components of cell walls, are present in lower amounts. The significant difference concerning the polysaccharide composition of musts and wines is discussed. Furthermore total polysaccharides were separated in calcium- and ethanol-precipitated polysaccharides. Their respective glycosyl-composition analyses, determined by GC of their per-<i>O</i>-trimethylsilylated methyl glycosides, revealed that neutral "gums" and acidic "pectins" could not be separated by specific precipitation.

The contribution of the O‐glycosylated protein Pir2p/Hsp150 to the construction of the yeast cell wall in wild‐type cells and β1,6‐glucan‐deficient mutants

The cell wall of yeast contains a major structural unit, consisting of a cell wall protein (CWP) attached via a glycosylphosphatidylinositol (GPI)-derived structure to beta 1,6-glucan, which is linked in turn to beta 1, 3-glucan. When isolated cells walls were digested with beta 1,6-glucanase, 16% of all CWPs remained insoluble, suggesting an alternative linkage between CWPs and structural cell wall components that does not involve beta 1,6-glucan. The beta 1,6-glucanase-resistant protein fraction contained the recently identified GPI-lacking, O-glycosylated Pir-CWPs, including Pir2p/Hsp150. Evidence is presented that Pir2p/Hsp150 is attached to beta 1,3-glucan through an alkali-sensitive linkage, without beta 1,6-glucan as an interconnecting moiety. In beta 1,6-glucan-deficient mutants, the beta 1,6-glucanase-resistant protein fraction increased from 16% to over 80%. This was accompanied by increased incorporation of Pir2p/Hsp150. It is argued that this is part of a more general compensatory mechanism in response to cell wall weakening caused by low levels of beta 1,6-glucan.

Bacterial Cell Wall Polymer-Induced Granulomatous Inflammation

Local or systemic injection of peptidoglycan-polysaccharide polymers, which are the primary structural components of cell walls of nearly all bacteria, leads to acute inflammation, which can develop into chronic, spontaneously relapsing, granulomatous inflammation in a number of organs. Evolution into chronic granulomatous inflammation is dependent upon persistence of poorly biodegradable cell wall polymers within tissues, genetically determined host susceptibility, and generation of a T-lymphocyte-mediated immune response. Intraperitoneal injection of peptidoglycan-polysaccharide fragments from group A streptococci or selected intestinal bacteria into susceptible Lewis rats leads to chronic, spontaneously reactivating erosive arthritis and hepatic granulomas. Subserosal (intramural) injection of poorly biodegradable cell wall fragments into the distal intestine of Lewis rats induces chronic, spontaneously relapsing granulomatous enterocolitis with associated arthritis, hepatic granulomas, anemia, and leukocytosis. Chronic inflammation does not occur in T-lymphocyte-deficient rats and is prevented by cyclosporin-A therapy and degradation of peptidoglycan by the muralytic enzyme, mutanolysin. Moreover, resistant Buffalo and Fischer F344rats, the latter sharing identical MHC antigens with Lewis rats, develop only acute inflammation with no chronic granulomatous response. Peptidoglycan-polysaccharide polymers activate almost every limb of the inflammatory response. Blockade of specific pathways suggests that interleukin-1, transforming growth factor-β, plasma kallikrein, and T lymphocytes are dominant mediators of peptidoglycan-polysaccharide-induced arthritis, hepatic granulomas, and enterocolitis. Because of the similarity of immune mechanisms of these rat models to human disease, bacterial cell wall-induced inflammation provides unique opportunities to study pathogenic mechanisms of granuloma formation in response to ubiquitous microbial agents and to test novel therapeutic agents.

Wood contains a cell-wall structural protein.

A pine extensin-like protein (PELP) has been localized in metabolically active cells of differentiating xylem and in mature wood of loblolly pine (Pinus taeda L.). This proline-rich glycosylated protein was purified from cell walls of differentiating xylem by differential solubility and gel electrophoresis. Polyclonal rabbit antibodies were raised against the deglycosylated purified protein (dPELP) and purified antibody was used for immunolocalization. Immunogold and alkaline phosphatase secondary antibody staining both show antigen in secondary cell walls of earlywood and less staining in latewood. Immunoassays of milled dry wood were developed and used to show increased availability of antigen after hydrogen fluoride or cellulase treatment and decreased antigen after chlorite treatment. The specificity of the antigen-antibody reaction was confirmed by competition assays and by preadsorption of antibody to the purified protein. We propose that extensin-like protein is present in xylem cell walls during lignification and that the protein remains as a structural component of cell walls in wood for many years after xylogenesis. We suggest that such structural proteins play important roles in the differentiation of xylem and thereby could affect the properties of wood.

Characterization of structural component of cell walls of alkalophilic strain of Bacillus sp. C-125. Preparation of poly(γ-l-glutamate) from cell wall component

The cell wall of an alkalophilic strain of Bacillus sp. C-125 is composed of A1 gamma-peptidoglycan, a teichuronic acid and an unknown acidic polymer composed of glutamic acid and glucuronic acid, of which the molar ratio is approx. 4-5:1. Poly(gamma-L-glutamate) was prepared from the acidic polymer by removal of almost all of the glucuronic residues with trifluoromethanesulphonic acid treatment and purified chromatographically. The Mr of the polyglutamate preparation was estimated to be 14,000 by gel chromatography, or 43,000 on the basis of the content of N-terminal acid residues. The acidic polymer found in the cell wall of the organism was concluded to be a polyglutamate substituted with (oligo)glucuronic acid residues or a complex composed of two kinds of polymers (polyglutamate and polyglucuronate).

Cell wall glycoproteins: structure and function.

Hydroxyproline-rich glycoproteins form an important, but little understood, structural component of most cell walls. Their occurrence, chemistry, synthesis, secretion, cross-linking and functions in higher plant cell walls will be briefly reviewed. Similar molecules also occur in other groups of plants; in particular, in the algae. In many of these they form highly ordered cell surface arrays, and we have studied these by high-resolution electron microscopy and computer image reconstruction. Some resulting three-dimensional models of these are presented. One particular glycoprotein, the major structural component of the cell wall of Chlamydomonas reinhardii, has been investigated in some detail. The chemistry and structure of this glycoprotein, which we have called volvin, has been studied and a family of monoclonal antibodies has been raised against it. Some of these antibodies appear to be specific to oligosaccharide side-chains and allow the localization of these substituents and their sites of synthesis. Immunofluorescence studies have shown that the expression of some of these antigenic determinants is developmentally regulated or cell-cycle-dependent. Immunogold labelling of thin sections has enabled the sites of synthesis and the method of secretion to be determined. These results will be discussed in the context of other cell wall glycoproteins, their relation to other glycoproteins, such as the mating agglutinin, and to their possible functions.

Enzymes of the yeast lytic system produced byArthrobacter GJM-1 bacterium and their role in the lysis of yeast cell walls

Yeast lytic system produced by Arthrobacter GJM-1 bacterium during growth on baker's yeast cell walls contains a complete set of enzymes which can hydrolyze all structural components of cell walls of Saccharomyces cerevisiae. Chromatographic fractionation of the lytic system showed the presence of two types of endo-beta-1,3-glucanase. Rapid lysis of isolated cell walls of yeast was induced only by endo-beta-1,3-glucanase exhibiting high affinity to insoluble beta-1,3-glucans and releasing laminaripentaose as the main product of hydrolysis of beta-1,3-glucans. This enzyme was able to lyse intact cells of S. cerevisiae only in the presence of an additional factor present in the Arthrobacter GJM-1 lytic system, which was identified as an alkaline protease. This enzyme possesses the lowest molecular weight among other identified enzyme components present in the lytic system. Its role in the solubilization of yeast cell walls from the outer surface by endo-beta-1,3-glucanase could be substituted by preincubation of cells with Pronase or by allowing the glucanase to act on cells in the presence of thiol reagents. The mechanism of lysis of intact cells and isolated cell walls by the enzymes of Arthrobacter GJM-1 is discussed in the light of the present conception of yeast cell wall structure.

Enzymes of the yeast lytic system produced by Arthrobacter GJM‐1 bacterium and their role in the lysis of yeast cell walls

AbstractYeast lytic system produced by Arthrobacter GJM‐1 bacterium during growth on baker's yeast cell walls contains a complete set of enzymes which can hydrolyze all structural components of cell walls of Saccharomyces cerevisiae. Chromatographic fractionation of the lytic system showed the presence of two types of endo‐ß‐1,3‐glucanase. Rapid lysis of isolated cell walls of yeast was induced only by endo‐ß‐1,3‐glucanase exhibiting high affinity to insoluble ß‐1,3‐glucans and releasing laminaripentaose as the main product of hydrolysis of ß‐1,3‐glucans. This enzyme was able to lyse intact cells of S. cerevisiae only in the presence of an additional factor present in the Arthrobacter GJM‐1 lytic system, which was identified as an alkaline protease. This enzyme possesses the lowest molecular weight among other identified enzyme components present in the lytic system. Its role in the solubilization of yeast cell walls from the outer surface by endo‐ß‐1,3‐glucanase could be substituted by preincubation of cells with Pronase or by allowing the glucanase to act on cells in the presence of thiol reagents. The mechanism of lysis of intact cells and isolated cell walls by the enzymes of Arthrobacter GJM‐1 is discussed in the light of the present conception of yeast cell wall structure.