Deglycosylation studies on tracheal mucin glycoproteins

Woodward, Harold D.; Ringler, Nancy J.; Selvakumar, R; Simet, Ira M.; Bhavanandan, V. P.; Davidson, Eugene A.

doi:10.1021/bi00391a015

Cited by 53 publications

(30 citation statements)

References 34 publications

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“…In parallel experiments, all of the glycosidases gave optimal digestion results when bovine fetuin was used as a control glycoprotein (data not shown). These results indicated that, although likely glycosylated, rmS5D-SRCRB-HA is also relatively resistant to glycosidase treatment, as has often been reported for some heavily glycosylated high M r proteins (25,26).…”

Section: Glycosylation Studiessupporting

confidence: 71%

Molecular and Functional Characterization of Mouse S5D-SRCRB: A New Group B Member of the Scavenger Receptor Cysteine-Rich Superfamily

Miró-Julià

Roselló

Martínez

et al. 2011

The Journal of Immunology

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The scavenger receptor cysteine-rich superfamily (SRCR-SF) members are transmembrane and/or secreted receptors exhibiting one or several repeats of a cysteine-rich protein module of ∼100 aa, named scavenger receptor cysteine-rich (SRCR). Two types of SRCR domains (A or B) have been reported, which differ in the number of coding exons and intradomain cysteines. Although no unifying function has been reported for SRCR-SF members, recognition of pathogen-associated molecular patterns (PAMPs) was recently shown for some of them. In this article, we report the structural and functional characterization of mouse S5D-SRCRB, a new group B member of the SRCR-SF. The s5d-srcrb gene maps at mouse chromosome 7 and encompasses 14 exons extending over 15 kb. The longest cDNA sequence found is 4286 bp in length and encodes a mature protein of 1371 aa, with a predicted Mr of 144.6 kDa. Using an episomal mammalian-expression system, a glycosylated soluble recombinant form >200 kDa was obtained and used as immunogen for the generation of specific rat mAbs. Subsequent immunohistochemical and real-time PCR analysis showed significant S5D-SRCRB expression in murine genitourinary and digestive tracts. S5D-SRCRB was shown to bind endogenous extracellular matrix proteins (laminin and galectin-1), as well as PAMPs present on Gram-positive and Gram-negative bacteria and fungi. PAMP binding by S5D-SRCRB induced microbial aggregation and subsequent inhibition of PAMP-induced cytokine release. These abilities suggest that S5D-SRCRB might play a role in the innate defense and homeostasis of certain specialized epithelial surfaces.

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Section: Glycosylation Studiessupporting

confidence: 71%

Molecular and Functional Characterization of Mouse S5D-SRCRB: A New Group B Member of the Scavenger Receptor Cysteine-Rich Superfamily

Miró-Julià

Roselló

Martínez

et al. 2011

The Journal of Immunology

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“…TFMSA nonselectively cleaves N-and O-linked sugars from the glycoproteins and deglcosylation of glycoproteins by TFMSA has been proven to be useful because the peptide backbone is left intact after treatment (22,23). Lewis rats immunized with CI-␣2 (22 kDa) after treatment with TFMSA (n ϭ 12) did not develop anterior uveitis (Fig.…”

Section: Posttranslational Modification Of the Ag Bestows Uveitogenicitymentioning

confidence: 99%

Type I Collagen Is the Autoantigen in Experimental Autoimmune Anterior Uveitis

Bora

Sohn

Kang

et al. 2004

The Journal of Immunology

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This study was undertaken to identify and characterize the Ag responsible for the induction of experimental autoimmune anterior uveitis (EAAU). Melanin-associated Ag isolated from bovine iris and ciliary body was digested with the proteolytic enzyme V8 protease to solubilize the proteins and the pathogenic protein was purified to homogeneity. Lewis rats were sensitized to various fractions and investigated for the development of anterior uveitis and an immune response to the purified Ag. The uveitogenic Ag had a mass of 22 kDa (SDS-PAGE) and an isoelectric point of 6.75. The N-terminal amino acid sequence of this protein demonstrated 100% homology with the bovine type I collagen α-2 chain starting from amino acid 385 and will be referred to as CI-α2 (22 kDa). Animals immunized with bovine CI-α2 (22 kDa) developed both cellular and humoral immunity to the Ag. They developed anterior uveitis only if the CI-α2 chain underwent proteolysis and if the bound carbohydrates were intact. EAAU induced by CI-α2 (22 kDa) can be adoptively transferred to naive syngenic rats by primed CD4+ T cells. EAAU could not be induced by the adoptive transfer of sera obtained from animals immunized with CI-α2 (22 kDa). The α-1 and α-2 chains (intact or proteolytically cleaved) of type I collagen from calfskin were not pathogenic. Although human anterior uveitis has been historically characterized as a collagen disease, this is first time collagen has been directly identified as the target autoantigen in uveitis.

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“…Chromosome mapping shows the two genes are located on different chromosomes, supporting a one-organ-one-gene hypothesis. On the other hand, a comparison of amino acid composition between airway and intestinal mucins (2,7,8) revealed a high degree of similarity. Furthermore, recent data showed that antibodies raised against the core protein ofbronchial mucin cross-reacted with mucin in intestinal goblet cells, strengthening the possibility of close structural homology between mucins from the two organs (13).…”

Section: Introductionmentioning

confidence: 99%

“…Biochemical analysis of purified mucins from various human tissues shows that they share several general properties: high molecular weight (> 106 D), high carbohydrate content (70-80%), and oligosaccharides consisting of N-acetyl galactosamine, galactose, fucose, sialic acid, and N-acetyl glucosamine (4). The latter are joined by O-glycosidic linkages to the core protein or apomucin, which is typically rich in hydroxyamino acids (2,(5)(6)(7). However, despite these similarities, extensive microheterogeneity exists, with mucins from various sources showing different stoichiometry of amino acids and sugars (compare 5, 7-9).…”

Section: Introductionmentioning

confidence: 99%

Human bronchus and intestine express the same mucin gene.

Jany¹,

Gallup²,

Yan³

et al. 1991

J. Clin. Invest.

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The amino acid and sugar composition of mucins from various organs is similar but not identical. This could arise by one or more of the following: organ-specific processing of a single core protein, organ-specific splicing of a single mucin mRNA, or organ-specific expression of various mucin genes. To begin to investigate the source ofthis variability, we examined (a) immunological cross-reactivity and (b) cDNA cross-hybridization, among several mucin-secreting organs of the human body. Peptide-directed antibodies raised against both nondeglycosylated (LS) and deglycosylated (HFB) intestinal mucin strongly stained mucous cells in the bronchial epithelium and submucosal glands, indicating homology between mucins of the bronchus and intestine at the peptide level. By screening a bronchus cDNA library with an intestinal mucin cDNA, SMUC-41, we isolated a bronchus mucin cDNA, HAM-1. This cDNA is 96% homologous to the first repeat of SMUC-41. HAM-I hybridized to restriction fragments of human genomic DNA identical to those hybridizing to SMUC-41 on Southern blots. SMUC-41 also hybridized to polydisperse transcripts in the bronchus, cervix, gall bladder, and mammary gland, indicating mucin homology among all these organs at the RNA level. We conclude that the bronchus and intestine express a common mucin gene, which is likely co-expressed by at least several other mucin-secreting organs. (J. Clin. Invest. 1991. 87:7742.)

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Deglycosylation studies on tracheal mucin glycoproteins

Cited by 53 publications

References 34 publications

Molecular and Functional Characterization of Mouse S5D-SRCRB: A New Group B Member of the Scavenger Receptor Cysteine-Rich Superfamily

Molecular and Functional Characterization of Mouse S5D-SRCRB: A New Group B Member of the Scavenger Receptor Cysteine-Rich Superfamily

Type I Collagen Is the Autoantigen in Experimental Autoimmune Anterior Uveitis

Human bronchus and intestine express the same mucin gene.

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