Your friend has discovered a protein that they suspect is glycosylated. They decide to perform a series of tests to determine the nature of the oligosaccharide. Assuming/Knowing the following: Fucose molecular weight is approximately 164 Galactose molecular weight is approximately 180 GalNAc molecular weight is approximately 221 Mannose molecular weight is approximately 180 Sialic Acid molecular weight is approximately 309 Neuraminidase cleaves before a Sialic Acid (Sialic Acid and anything after leaves the protein) Beta-galactosidase cleaves after a Galactose (Galactose and anything before remains on the protein) Peanut Agglutinin binds to GalNAc Concanavalin A binds to Mannose Hemagglutinin binds to Sialic Acid Based on preliminary results, they suspect that the oligosaccharide is 7 glycosides long and weighs a total of 1414 g/mol They carried out the following experiments: they treated the protein with either a glycosidase or a lectin, and then pelleted the protein using ammonium sulfate purification. They then used various means to measure the mass of the released (not protein bound) sugars and glycosides. They also have a way of detecting where a particular lectin binds, or if it is still present in the supernatant (or both). Here are the results: Treatment Released Oligosaccharides Peanut Agglutinin Concanavalin A Hemagglutinin Beta-Galactosidase 890 and 164 Soluble Soluble and Pellet Soluble Neuraminidase 833 Pellet Soluble and Pellet Soluble Beta-Galactosidase & Neuraminidase 669, 221, and 164 Soluble Soluble and Pellet Soluble What is the sequence of their oligosaccharide? Explain your answer!
Nucleotides
It is an organic molecule made up of three basic components- a nitrogenous base, phosphate,and pentose sugar. The nucleotides are important for metabolic reactions andthe formation of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid).
Nucleic Acids
Nucleic acids are essential biomolecules present in prokaryotic and eukaryotic cells and viruses. They carry the genetic information for the synthesis of proteins and cellular replication. The nucleic acids are of two types: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). The structure of all proteins and ultimately every biomolecule and cellular component is a product of information encoded in the sequence of nucleic acids. Parts of a DNA molecule containing the information needed to synthesize a protein or an RNA are genes. Nucleic acids can store and transmit genetic information from one generation to the next, fundamental to any life form.
Your friend has discovered a protein that they suspect is glycosylated. They decide to perform a series of tests to determine the nature of the oligosaccharide.
Assuming/Knowing the following:
- Fucose molecular weight is approximately 164
- Galactose molecular weight is approximately 180
- GalNAc molecular weight is approximately 221
- Mannose molecular weight is approximately 180
- Sialic Acid molecular weight is approximately 309
- Neuraminidase cleaves before a Sialic Acid (Sialic Acid and anything after leaves the protein)
- Beta-galactosidase cleaves after a Galactose (Galactose and anything before remains on the protein)
- Peanut Agglutinin binds to GalNAc
- Concanavalin A binds to Mannose
- Hemagglutinin binds to Sialic Acid
Based on preliminary results, they suspect that the oligosaccharide is 7 glycosides long and weighs a total of 1414 g/mol
They carried out the following experiments: they treated the protein with either a glycosidase or a lectin, and then pelleted the protein using ammonium sulfate purification. They then used various means to measure the mass of the released (not protein bound) sugars and glycosides. They also have a way of detecting where a particular lectin binds, or if it is still present in the supernatant (or both).
Here are the results:
| Treatment | Released Oligosaccharides | Peanut Agglutinin | Concanavalin A | Hemagglutinin |
| Beta-Galactosidase | 890 and 164 | Soluble | Soluble and Pellet | Soluble |
| Neuraminidase | 833 | Pellet | Soluble and Pellet | Soluble |
| Beta-Galactosidase & Neuraminidase | 669, 221, and 164 | Soluble | Soluble and Pellet | Soluble |
What is the sequence of their oligosaccharide? Explain your answer!
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