Human Natural Killer-1 Sulfotransferase (HNK-1ST)-induced Sulfate Transfer Regulates Laminin-binding Glycans on α-Dystroglycan

Nakagawa, Naoki; Manya, Hiroshi; Toda, Tatsushi; Endo, Tamao; Oka, Shogo

doi:10.1074/jbc.m112.363036

Cited by 30 publications

(30 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Western blotting. Western blots were performed according to methods in previous studies 29,30 . At 72 h post transfection, the EPO and sFγRIII recombinant proteins were purified with anti-Flag M2 Affinity Gel (Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor

et al. 2020

View full text Add to dashboard Cite

MCFD2 and ERGIC-53, which are the products of causative genes of combined factor V and factor VIII deficiency, form a cargo receptor complex responsible for intracellular transport of these coagulation factors in the early secretory pathway. In this study, using an NMR technique, we successfully identified an MCFD2-binding segment from factor VIII composed of a 10 amino acid sequence that enhances its secretion. This prompted us to examine possible effects of attaching this sequence to recombinant glycoproteins on their secretion. We found that the secretion level of recombinant erythropoietin was significantly increased simply by tagging it with the passport sequence. Our findings not only provide molecular basis for the intracellular trafficking of coagulation factors and their genetic deficiency but also offer a potentially useful tool for increasing the production yields of recombinant glycoproteins of biopharmaceutical interest.

show abstract

Section: Methodsmentioning

confidence: 99%

Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor

et al. 2020

View full text Add to dashboard Cite

show abstract

“…cDNA Construction-For the expression of Fc-fused ␣DG recombinant proteins, amino acid substitutions and deletion mutants of ␣DG (␣DG373(T322R)) ( Fig. 1) were made by standard PCR and genetic engineering techniques using the expression plasmids as constructed previously (7,16). For Flag-tagged expression vectors of human FKTN, FKRP, TMEM5, and ISPD, the encoding sequence of individual open reading frames was amplified by PCR and cloned into pCMV14 -3ϫFLAG vector (Sigma).…”

mentioning

confidence: 99%

Direct Mapping of Additional Modifications on Phosphorylated O-glycans of α-Dystroglycan by Mass Spectrometry Analysis in Conjunction with Knocking Out of Causative Genes for Dystroglycanopathy

Yagi

Kuo

Obayashi

et al. 2016

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

“…7 Loss of the a-DG subunit in melanoma is likely to be caused by several post-translational mechanisms, such as an altered glycosylation pattern and/or proteolytic degradation of the membrane complex. 8 Loss of a-DG function results in a disruption of cell-to-ECM interactions and might promote tumour invasion and metastasis. However, the observed expression of a-DG on cells with spindle-cell like morphology in melanoma metastasis suggests that its restoration could favour the implant of cells in metastatic sites, as previously reported in other cancers.…”

Section: Loss Of Alpha-dystroglycan Expression In Cutaneous Melanocytmentioning

confidence: 99%

Loss of alpha‐dystroglycan expression in cutaneous melanocytic lesions

Garcovich

Migaldi

Bonetti

et al. 2015

Acad Dermatol Venereol

View full text Add to dashboard Cite

The dystroglycan complex (DG), originally characterized in muscle and in genetic muscle diseases, links the epithelial cell cytoskeleton to the basement membrane.[1] DG has been shown to be involved in skin morphogenesis and in epithelial carcinogenesis, modulating cell differentiation and adhesion, assembly of the epithelial basement membrane and interactions with the extracellular matrix (ECM).[2] DG comprises an alpha (α-DG) and a beta (β-DG)-subunits, with the α-DG subunit being the extracellular, functional part of the complex and binding several ECM components. Loss of α-DG function has been reported in several epithelial- and neural-derived tumours and correlated with tumour grading, progression and clinical outcome.[3] In the skin, DG is localized at the dermo-epidermal junction and is produced by epidermal keratinocytes and dermal fibroblasts.[4] Tissue expression of the α-DG subunit has not been previously characterized in cutaneous melanocytic nevi and in malignant melanoma. In this study, expression of the DG complex was analysed by Western-blot in cell extracts from human normal melanocytes (hMel) and three melanoma cell lines (BLM, M14, IF6). Furthermore, tissue expression of α-DG was assessed by immunohistochemistry in a panel of archival melanocytic lesions, including melanocytic nevi (n = 20), cutaneous melanoma (n = 51) and melanoma metastases (n = 53). A polyclonal antibody to alpha-DG (clone VIA4-1) from Upstate Biotechnology and a monoclonal antibody to β-DG (clone 43DAG/8D5) from Novocastra were used for the analyses. β-DG molecule was detectable in both melanocytes and melanoma cells displaying a higher expression in IF6 melanoma cells compared to normal melanocytes. On the other hand, a decreased expression of α-DG was observed in all three melanoma cell lines compared to normal melanocytes (Fig. 1). At the tissue level, in benign melanocytic nevi α-DG was expressed by basal melanocytes at the dermo-epidermal junction and by epithelioid melanocytes, organized in nests, in the superficial dermis (Fig. 2a,b). In primary cutaneous melanoma, α-DG expression appeared to be lost in almost all cases. Loss of α-DG expression in melanoma was observed at all Clark invasion levels, both in the epidermis and dermo-epidermal junction as well as in the superficial to deep dermis (Fig. 2c–e). In melanoma metastases, expression of α-DG was evident in 20.8% of cases, being mainly focal and limited to spindle-cell like melanoma cells (Fig. 2f). Previous studies have only characterized the in-vitro expression of DG subunits by melanocytes and melanoma cells, as well as tissue expression of β-DG in melanocytic nevi.[5, 6] We report a differential pattern of α-DG expression between benign and malignant melanocytic tumours, suggesting a peculiar role of the DG complex in melanocyte biology and melanomagenesis. In benign melanocytic nevi, α-DG displayed a consistent expression pattern at the dermo-epidermal junction, where interactions with basal keratinocytes and ECM components are prominent in the epidermal me...

show abstract

Human Natural Killer-1 Sulfotransferase (HNK-1ST)-induced Sulfate Transfer Regulates Laminin-binding Glycans on α-Dystroglycan

Cited by 30 publications

References 53 publications

Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor

Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor

Direct Mapping of Additional Modifications on Phosphorylated O-glycans of α-Dystroglycan by Mass Spectrometry Analysis in Conjunction with Knocking Out of Causative Genes for Dystroglycanopathy

Loss of alpha‐dystroglycan expression in cutaneous melanocytic lesions

Contact Info

Product

Resources

About