中山大学蛋白质组学技术分析.docx

I.Introduction Structure ?Building blocks——amino acids ?Peptide bond and peptides ?Primary structure of protein ?The three-dimensional structure of proteins ?Getting protein structure ?Integral membrane proteins ?Relationship between structure and function of proteins Function ?Principle in general ?Protein folding, misfolding and disease Regulation? * Expression * TransportationLocalization * Modification(Phosphorylation, glycosylation, methylation, acetylation, and ubiquitin) * Interaction * Degradation Levels of structure 1°: Amino acid residue sequence 2°: Structural elements 3°: Specific 3-dimensional structure 4°: Arrangement of subunits in multi-subunit protein Note? ? ?16% N ? Met for metabolic labelling ? Trp, Tyr, and to a lesser extent Phe, absorb ultraviolet light. This accounts for the characteristic strong absorbance of ? light by proteins at a wavelength of 280 nm. ? Count the amino acid numbers in protein.?138???? 128???? 110 ? symbol ?Nonstandard amino acids?(hydroxyproline,hydroxylysine) ?Some additional amino acids ?selenocysteine(硒半胱氨酸)andpyrrolysine(吡咯赖氨酸) Chemical synthesis of peptides ????? Liquid (Solution ) phase peptide synthesis ????? Solid phase peptide synthesis（SPPS） ????? Pre-treatment of amino acids ????? Formation of peptide bond ????? Deprotect, hydrolyze and purification Difference between biological?and chemical synthesis of peptides ? Template ? Orientation 生物NC 化学CN ? Efficiency (A protein with 100 amino acids : Chemical synthesis--about 4 days; Biological process--about 5 seconds in E. coli) ? Condition ? size Sequence ? Sanger 1953 胰岛素 ? 反推DNA（简并性） ? 抗原抗体 核糖体 测mRNA ? 测一部分比对数据库 Motifs(units) structure motifs 1) Helix-loop-helix2 αhelices joined by bridge? 2) βhairpin2 adjacent antiparallel βstrands connected by a β-turn. 3) β? α? βloopAn αhelix serves as the connection between 2 parallel βstrands. function motifs ? ?Thr-Gly-Tyr,Thr-Pro-Tyr motif Domains? 1) All αdomainsrepeat of helix-loop-helix motif 2) All βdomainsrepeat of β