Lectin Microarray Profiling and Relative Quantification of Glycome Associated With Proteins of Neonatalwtandrd1Mice Retinae

Abstract

Citation: Ahuja S. Lectin microarray profiling and relative quantification of glycome associated with proteins of neonatal wt and rd1 mice retinae. Invest Ophthalmol Vis Sci. 2013;54:3272-3280. DOI:10.1167/ iovs.12-11363 PURPOSE. Progressive dynamic, relative quantitative changes were compared in glycans associated with retinal proteins of wild type (wt) and retinal degeneration 1 (rd1) mice during neonatal development and degeneration of retinae. METHODS.Proteins extracted from retinae of postnatal days 2 (PN2), PN7, and PN14 wt and rd1 mice were labeled with Cy3-fluorescent dye. Glycome of these proteins was quantified relatively by lectin microarray technique. Net fluorescence emitted by individual complexes formed between 45 lectins and Cy3-labeled proteins was measured by evanescent-field fluorescence-assisted microarray reader.RESULTS. GlcNAcb1-oligomer and high-mannose/Mana1-6Man were major glycans associated with the proteins of PN2, PN7, and PN14 wt and rd1 mice retinae. Gal/GalNAc/Man 3 -core-bi-/ tri-antennary-complex, Sia2-3Galb1-4GlcNAc, and high-mannose glycans were conjugated mainly to proteins from PN7 rd1 and PN14 wt retinae, respectively. With increasing neonatal age, mannosylated, GlcNAcb, and sialylated (minor component) glycans were increased, and fucosylated GlcNAc/Galb glycans were decreased significantly in wt retinal proteins. This trend was less evident in PN14 rd1 retinal proteins. Mouse retina was almost devoid of Siaa2-6 (except WGA bound Sia), Fuca1-2, and Gal/GalNAc-containing glycans. STL reacting GlcNAc oligomers were high in PN2 rd1 retinae.CONCLUSIONS. Quantitative dynamic, relative variation in high-mannose and GlcNAc glycans, Siaa2-3Galb1-4GlcNAc associated with proteins from PN2, PN7, and PN14 wt and rd1 mice retinae suggested that these glycans participate in retinal development and degeneration, and may be used as markers for retinal electrophysiologic integrity during transplantation/therapy studies; Siaa2-3Galb1-4GlcNAc-specific Agrocybe cylindracea lectin and other lectins may be used to enrich/purify retinal ribbon synapse glycoproteins and other glycoproteins including rhodopsin. Further investigations are required.

Binding of the SARS-CoV-2 spike protein to glycans

“…The microarray experiment was performed using a previously established standard protocol. 29,39,40 Briefly, the proteins were labeled with Cy3 fluorescent dye and incubated with the microarray at different concentrations. After washing away the unbound HS molecules, a highly sensitive fluorescence method was used to detect the binding of the SARS-CoV-2-RBD to HS.…”

Comprehensive profiling of accessible surface glycans of mammalian sperm using a lectin microarray

“…The lectin microarrays have been applied for fast and high-throughput glycan profiling and comparison for a variety of samples, e.g. , glycoprotein [22-26], cell lysates [27-29], clinical specimens [30,31], mammalian cells [20,32-34], bacteria [17,35] and virus [36]. Using the lectin microarray, the disease specific or cell type specific lectins could be easily identified.…”

Determination of Dynamic Changes in the Nature and Biosynthesis of Glycome ofwtandrd1Mice Retinae by Lectin Microarray Analysis

“…The dynamic changes in mice retinal glycans have been reported previously. 1 In the available retinal Proteomics studies, 2 the glycoproteome was not characterized. Glycans and glycan levels determined by lectin microarray (see fig.…”

“…The categorization of glycans, with respective GT/G, lectins (abbreviated) binding specificity, and significance of differences, presents another facet of the lectin microarray data shown in figure 4 from a prior report by Ahuja. 1 Glycan functions are from the literature.…”