N-glycoproteomic and proteomic alterations in SRD5A3-deficient fibroblasts.

IF 3.4 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Glycobiology Pub Date : 2024-09-30 DOI:10.1093/glycob/cwae076
Kishore Garapati, Wasantha Ranatunga, Neha Joshi, Rohit Budhraja, Saniha Sabu, Kristin A Kantautas, Graeme Preston, Ethan O Perlstein, Tamas Kozicz, Eva Morava, Akhilesh Pandey
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Abstract

SRD5A3-CDG is a congenital disorder of glycosylation (CDG) resulting from pathogenic variants in SRD5A3 and follows an autosomal recessive inheritance pattern. The enzyme encoded by SRD5A3, polyprenal reductase, plays a crucial role in synthesizing lipid precursors essential for N-linked glycosylation. Despite insights from functional studies into its enzymatic function, there remains a gap in understanding global changes in patient cells. We sought to identify N-glycoproteomic and proteomic signatures specific to SRD5A3-CDG, potentially aiding in biomarker discovery and advancing our understanding of disease mechanisms. Using tandem mass tag (TMT)-based relative quantitation, we analyzed fibroblasts derived from five patients along with control fibroblasts. N-glycoproteomics analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) identified 3,047 glycopeptides with 544 unique N-glycosylation sites from 276 glycoproteins. Of these, 418 glycopeptides showed statistically significant changes with 379 glycopeptides decreased (P < 0.05) in SRD5A3-CDG patient-derived samples. These included high mannose, complex and hybrid glycan-bearing glycopeptides. High mannose glycopeptides from protocadherin Fat 4 and integrin alpha-11 and complex glycopeptides from CD55 were among the most significantly decreased glycopeptides. Proteomics analysis led to the identification of 5,933 proteins, of which 873 proteins showed statistically significant changes. Decreased proteins included cell surface glycoproteins, various mitochondrial protein populations and proteins involved in the N-glycosylation pathway. Lysosomal proteins such as N-acetylglucosamine-6-sulfatase and procathepsin-L also showed reduced levels of phosphorylated mannose-containing glycopeptides. Our findings point to disruptions in glycosylation pathways as well as energy metabolism and lysosomal functions in SRD5A3-CDG, providing clues to improved understanding and management of patients with this disorder.

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SRD5A3 缺陷成纤维细胞中 N-糖蛋白组和蛋白质组的改变
SRD5A3-CDG是一种先天性糖基化障碍(CDG),由SRD5A3的致病变体引起,为常染色体隐性遗传。SRD5A3编码的酶--多聚戊烯醛还原酶在合成N-连接糖基化所必需的脂质前体方面起着至关重要的作用。尽管功能性研究对其酶功能有了深入了解,但在理解患者细胞的整体变化方面仍存在差距。我们试图鉴定 SRD5A3-CDG 特异的 N-糖基化蛋白质组和蛋白质组特征,这可能有助于生物标记物的发现并增进我们对疾病机制的了解。我们使用基于串联质量标记(TMT)的相对定量法分析了来自五名患者的成纤维细胞和对照成纤维细胞。通过液相色谱-串联质谱(LC-MS/MS)进行N-糖蛋白组学分析,从276种糖蛋白中鉴定出3,047个糖肽和544个独特的N-糖基化位点。其中,418 个糖肽发生了统计学意义上的显著变化,379 个糖肽减少(P<0.05)。
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来源期刊
Glycobiology
Glycobiology 生物-生化与分子生物学
CiteScore
7.50
自引率
4.70%
发文量
73
审稿时长
3 months
期刊介绍: Established as the leading journal in the field, Glycobiology provides a unique forum dedicated to research into the biological functions of glycans, including glycoproteins, glycolipids, proteoglycans and free oligosaccharides, and on proteins that specifically interact with glycans (including lectins, glycosyltransferases, and glycosidases). Glycobiology is essential reading for researchers in biomedicine, basic science, and the biotechnology industries. By providing a single forum, the journal aims to improve communication between glycobiologists working in different disciplines and to increase the overall visibility of the field.
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