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Method Article
A protocol for high-throughput screening, analysis and validation of protein glycopatterns
Zheng Li
Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an, 710069, PR China.
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
In the past two decades, one of the main ways to investigate glycosylation was using lectins. Lectins are carbohydrate-binding proteins that discriminate glycans based on subtle differences in structure. Recently, with the advent of high-throughput glycomic techniques, such as lectin microarray which is a system biology approach is utilized in order to draw general conclusions of protein glycosylation about a biological sample. The display of the lectins in a microarray enables the multiple and distinct binding glycopatterns to be observed simultaneously, providing information on the carbohydrate composition of the samples. The lectin blotting can be used to verify rapidly the target glycopatterns of glycoproteins by comparison with a reference SDS-PAGE protein map, and obtain reliable and reproducible results according to the lectin microarrays. Here, the lectin microarrays are used to probe the relative expression levels of terminal α2-3/6-linked sialic acids in human saliva from type 2 diabetes mellitus (T2DM). And, lectin blotting is used to confirm the different abundance of the terminal Siaα2-6Gal and Siaα2-3Gal between T2DM and healthy group according to the results of the lectin microarrays. In this protocol, we provide a detailed methodology of lectin microarray and lectin blotting for protein glycopattern analysis.
Keywords
Lectin microarrays, Lectin blotting, Saliva, glycoproteins, glycopattern
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Associated Publications
Avian Influenza Virus Infection Risk in Humans with Chronic Diseases
by Yaogang Zhong, Yannan Qin, Hanjie Yu, +13
Scientific Reports (20 March, 2015)
Age- and Sex-Associated Differences in the Glycopatterns of Human Salivary Glycoproteins and Their Roles against Influenza A Virus
by Yannan Qin, Yaogang Zhong, Minzhi Zhu, +7
Journal Of Proteome Research (21 March, 2015)
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This is a preprint. It has not completed peer review.
Method Article
A protocol for high-throughput screening, analysis and validation of protein glycopatterns
Zheng Li
Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an, 710069, PR China.
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 24 Mar, 2015
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
In the past two decades, one of the main ways to investigate glycosylation was using lectins. Lectins are carbohydrate-binding proteins that discriminate glycans based on subtle differences in structure. Recently, with the advent of high-throughput glycomic techniques, such as lectin microarray which is a system biology approach is utilized in order to draw general conclusions of protein glycosylation about a biological sample. The display of the lectins in a microarray enables the multiple and distinct binding glycopatterns to be observed simultaneously, providing information on the carbohydrate composition of the samples. The lectin blotting can be used to verify rapidly the target glycopatterns of glycoproteins by comparison with a reference SDS-PAGE protein map, and obtain reliable and reproducible results according to the lectin microarrays. Here, the lectin microarrays are used to probe the relative expression levels of terminal α2-3/6-linked sialic acids in human saliva from type 2 diabetes mellitus (T2DM). And, lectin blotting is used to confirm the different abundance of the terminal Siaα2-6Gal and Siaα2-3Gal between T2DM and healthy group according to the results of the lectin microarrays. In this protocol, we provide a detailed methodology of lectin microarray and lectin blotting for protein glycopattern analysis.
Figure 1
Associated Publications
Avian Influenza Virus Infection Risk in Humans with Chronic Diseases
by Yaogang Zhong, Yannan Qin, Hanjie Yu, +13
Scientific Reports (20 March, 2015)
Age- and Sex-Associated Differences in the Glycopatterns of Human Salivary Glycoproteins and Their Roles against Influenza A Virus
by Yannan Qin, Yaogang Zhong, Minzhi Zhu, +7
Journal Of Proteome Research (21 March, 2015)