A protocol for high-throughput screening, analysis and validation of protein glycopatterns
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
Posted 24 Mar, 2015
A protocol for high-throughput screening, analysis and validation of protein glycopatterns
Posted 24 Mar, 2015
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
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