生物化学资料：蛋白质一级结构.pptVIP

Structure of Proteins Primary Structure of proteins Whats the the primary stucture of proteins? primary structure of proteins overview Peptide bond AA coposition of a polypetide Sequencing of the peptide Peptide bond Peptide bond amide linkages betw. the α-COOH of one AA the α-NH2 of another NOT broken by conditions that denature proteins. nonenzymically hydrolyzed by a strong acid base at elevated temperatures. Naming the Peptide N-end C-end from: Each AA in a polypeptide is called a “residue” Suffixes(-ine,-an,ic or -ate) change to -yl,exept for C-terminal AA Naming the Peptide 谷胱甘肽：γ-谷氨酰-半胱氨酰-甘氨酸 Characteristics of the peptide bond Partial double bond character Nearly always in the trans configuration the atoms of the group, O=C-N-H, are fixed on the same plane Polarity:The –C=O -NH of the peptide bond uncharge but polar, are involved in H-bond Polarity of the peptide bond -C=O and -NH AA coposition of a polypetide Determination of the AA composition of a polypeptide Amino acid analyzer (阳离子交换层析技术） hydrolyz by strong acid at 110℃for 24hr ninhydrin（茚三酮） photometer（分光光度计） cation-exchange chromatography ninhydrin reaction Sequencing of the peptide Sequencing of the peptide from :N-end from :C-end DNA sequence Edman degradation Aminopeptidase Carboxypeptidase Edman degradation method Edman reagent: phenylisothiocyanate（PITC/PTC ,异硫氰酸苯酯） phenylthiohydantoin （PTH，苯乙内酰硫脲） resulting an instabilaty in the N-terminal peptide bond Label the N-terminal residue under mildly basic conditions Selective hydrolyzation,cycling, identification(HPLC) Repeat cycle(1cycle/hr) Aminopeptidase(氨肽酶): exopeptidase,cleavaging the peptide from its N-terminal end. According to the quantity and variety?of AA in a certain time to sequence the ptptide. Petidases method Carboxypeptidase(羧肽酶): exopeptidase,cleavaging the peptide from its C-terminal end. Cyanoen bromide(溴化氰)：cleave at Met residue Cleavage of the polypeptide into smaller fragments Sequencing