proteins_1_primary_structure__purificati

proteins_1_primary_structure__purificati - Ch Chemistry 237...

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Chemistry 237 roteins 1: Purification and Proteins 1: Purification and Primary Sequence Determination
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earning Goals Learning Goals earn the basics of protein purification Learn the basics of protein purification • Learn the techniques of protein equencing sequencing • Discuss some of the uses of sequences
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eptides and proteins Peptides and proteins • Linear sequence of amino acids. ne free N One free N terminus and one free C terminus. • Held together by peptide bonds • Sequences are ritten from the N written from the N to the C terminus.
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hy we bother Why we bother Proteins are the agents of biological function •E n z ymes - Ribonuclease • Regulatory proteins - Insulin • Transport proteins - Hemoglobin • Structural proteins - Collagen ontractile proteins - ctin, Myosin Contractile proteins Actin, Myosin • Exotic proteins - Antifreeze proteins in fish, green fluorescent protein p
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Why we bother Catalysis: –enolase (in the glycolytic pathway) –DNA polymerase (in DNA replication) Transport: –hemoglobin (transports O 2 in the blood) –lactose permease (transports lactose across the cell membrane) • Structure: ollagen (connective tissue) –collagen (connective tissue) –keratin (hair, nails, feathers, horns) • Motion: –myosin (muscle tissue) –actin (muscle tissue, cell motility)
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What to Study about Peptides nd Proteins? and Proteins? What is its sequence and composition? What is its three-dimensional structure? How does it find its native fold? How does it achieve its biochemical role? How is its function regulated? How does it interacts with other macromolecules? How is it related to other proteins? Where is it localized within the cell? What are its physico-chemical properties?
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Covalent Structures of Proteins Sequence determination techniques: 1) Gene sequence. Each amino acid is oded for by 3 nucleotides DNA coded for by 3 nucleotides. DNA sequence can be translated into protein equence. sequence. ) Chemical and enzymatic sequencing of 2) Chemical and enzymatic sequencing of purified protein. Mass spectrometry equencing. sequencing.
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rotein Purification Protein Purification now gene sequence. Know gene sequence. now protein sequence Know protein sequence. now size charge maybe something about the Know size, charge, maybe something about the structure and function. Design a purification scheme based on those roperties properties
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rotein Purification Protein Purification now protein function. Know protein function. on’t know protein sequence Don t know protein sequence. on’t know size charge maybe something about Don t know size, charge, maybe something about the structure Try different separation techniques and monitor rotein concentration and function protein concentration and function.
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rotein Purification Protein Purification eparation relies on differences in Separation relies on differences in physico-chemical properties ize Size – Charge – Affinity for a ligand – Solubility – Hydrophobicity – Thermal stability • Chromatography is commonly used for preparative separation
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This note was uploaded on 07/07/2010 for the course CHEM 237 taught by Professor Elisabethdaub during the Fall '10 term at Waterloo.

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proteins_1_primary_structure__purificati - Ch Chemistry 237...

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