Neurospora crassa family GH72 glucanosyltransferases function to crosslink cell wall glycoprotein N-linked galactomannan to cell wall lichenin

Kar, Bibekananda; Patel, Pavan; Ao, Jie; Free, Stephen J.

doi:10.1016/j.fgb.2018.11.007

Cited by 16 publications

(22 citation statements)

References 40 publications

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“…β-1,3-glucan is a long unbranched polysaccharide consisting of repeating β-1,3-glucose residues. It is the most abundant component of the vegetative cell wall, making up approximately 35% of the N. crassa cell wall mass (Kar et al, 2019). It is the major component of the cell walls found in almost all fungi.…”

Section: β-13-glucanmentioning

confidence: 99%

“…The linkage analysis of the N. crassa vegetative cell wall shows that between 15 and 20% of the glucoses in the wall have a 1,4 linkage and lichenin has been shown to be present as defined by a monoclonal antibody directed against lichenin (Ao and Free, 2017; Kar et al, 2019). Lichenin is defined as a polysaccharide with a repeating β-1,4-glucose-β-1,4-glucose-β-1,3-glucose trisaccharide (Perlin and Suzuki, 1962).…”

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

“…Lichenin has been shown to be located in the fungal cell wall and in the extracellular matrix formed by the ascomycete cells in the lichen (Honegger and Haisch, 2001). Based on this structure and assuming that all of the 1,4-linked glucose in the cell wall linkage analysis come from lichenin, lichenin would represent approximately 25% of the N. crassa vegetative cell wall mass (Kar et al, 2019). It has been shown that lichenin functions as the polysaccharide through which cell wall glycoproteins are cross-linked into the cell wall (Kar et al, 2019).…”

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

“…Based on this structure and assuming that all of the 1,4-linked glucose in the cell wall linkage analysis come from lichenin, lichenin would represent approximately 25% of the N. crassa vegetative cell wall mass (Kar et al, 2019). It has been shown that lichenin functions as the polysaccharide through which cell wall glycoproteins are cross-linked into the cell wall (Kar et al, 2019). Lichenin may also function to cross-link the β-1,3-glucan together into a matrix structure, but this has not been experimentally verified.…”

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

“…This demonstrates the important roles that N-linked oligosaccharides play in the process of protein folding, protein stability, and in the incorporation of glycoproteins into the cell wall. Glycan profiling and glycan linkage analysis of the N-linked glycans present on cell wall glycoproteins in the Δ och-1 mutant (which lacks the N-linked oligosaccharide-associated galactomannan described below) demonstrates that N. crassa glycoproteins have a typical 2 N -acetylglucosamine:9 mannose N-linked oligosaccharide (Deshpande et al, 2008; Kar et al, 2019). Trimming of some of the terminal mannoses occurs on most of the N-linked oligosaccharides and contributes to the heterogeneity seen in N-linked oligosaccharides (Kar et al, 2019).…”

Section: Glycoproteinsmentioning

confidence: 99%

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The Genetics and Biochemistry of Cell Wall Structure and Synthesis in Neurospora crassa, a Model Filamentous Fungus

Patel

Free

2019

Front. Microbiol.

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This review discusses the wealth of information available for the N. crassa cell wall. The basic organization and structure of the cell wall is presented and how the wall changes during the N. crassa life cycle is discussed. Over forty cell wall glycoproteins have been identified by proteomic analyses. Genetic and biochemical studies have identified many of the key enzymes needed for cell wall biogenesis, and the roles these enzymes play in cell wall biogenesis are discussed. The review includes a discussion of how the major cell wall components (chitin, β-1,3-glucan, mixed β-1,3-/ β-1,4- glucans, glycoproteins, and melanin) are synthesized and incorporated into the cell wall. We present a four-step model for how cell wall glycoproteins are covalently incorporated into the cell wall. In N. crassa, the covalent incorporation of cell wall glycoproteins into the wall occurs through a glycosidic linkage between lichenin (a mixed β-1,3-/β-1,4- glucan) and a “processed” galactomannan that has been attached to the glycoprotein N-linked oligosaccharides. The first step is the addition of the galactomannan to the N-linked oligosaccharide. Mutants affected in galactomannan formation are unable to incorporate glycoproteins into their cell walls. The second step is carried out by the enzymes from the GH76 family of α-1,6-mannanases, which cleave the galactomannan to generate a processed galactomannan. The model suggests that the third and fourth steps are carried out by members of the GH72 family of glucanosyltransferases. In the third step the glucanosyltransferases cleave lichenin and generate enzyme/substrate intermediates in which the lichenin is covalently attached to the active site of the glucanosyltransferases. In the final step, the glucanosyltransferases attach the lichenin onto the processed galactomannans, which creates new glycosidic bonds and effectively incorporates the glycoproteins into the cross-linked cell wall glucan/chitin matrix.

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Section: β-13-glucanmentioning

confidence: 99%

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

Section: Mixed β-13/β-14 Glucans (Lichenin)mentioning

confidence: 99%

Section: Glycoproteinsmentioning

confidence: 99%

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The Genetics and Biochemistry of Cell Wall Structure and Synthesis in Neurospora crassa, a Model Filamentous Fungus

Patel

Free

2019

Front. Microbiol.

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2019–2020

Harvey

2023

Mass Spectrometry Reviews

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This review is the tenth update of the original article published in 1999 on the application of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2020. Also included are papers that describe methods appropriate to analysis by MALDI, such as sample preparation techniques, even though the ionization method is not MALDI. The review is basically divided into three sections: (1) general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation, quantification and the use of arrays. (2) Applications to various structural types such as oligo‐ and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals, and (3) other areas such as medicine, industrial processes and glycan synthesis where MALDI is extensively used. Much of the material relating to applications is presented in tabular form. The reported work shows increasing use of incorporation of new techniques such as ion mobility and the enormous impact that MALDI imaging is having. MALDI, although invented nearly 40 years ago is still an ideal technique for carbohydrate analysis and advancements in the technique and range of applications show little sign of diminishing.

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Cell surface display of proteins on filamentous fungi

Urbar-Ulloa

Montaño-Silva

Ramírez-Pelayo

et al. 2019

Appl Microbiol Biotechnol

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Neurospora crassa family GH72 glucanosyltransferases function to crosslink cell wall glycoprotein N-linked galactomannan to cell wall lichenin

Cited by 16 publications

References 40 publications

The Genetics and Biochemistry of Cell Wall Structure and Synthesis in Neurospora crassa, a Model Filamentous Fungus

The Genetics and Biochemistry of Cell Wall Structure and Synthesis in Neurospora crassa, a Model Filamentous Fungus

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2019–2020

Cell surface display of proteins on filamentous fungi

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