无指盘臭蛙皮肤抗茵肽的分离纯化与氨基酸序列测定.pdfVIP

应用与环境生物学报 2007，13(6)：834～839 Chin JAppl Environ Biol=ISSN1006-687X 2007一l2-25 无指盘臭蛙皮肤抗茵肽的分离纯化与氨基酸序列测定术 刘炯宇 江建平 何开泽 谢锋 郑中华 (中国科学院成都生物研究所 成都 610041) 摘 要 在抗菌活性跟踪下对四川冕宁产无指盘臭蛙(Odorrana grahami)皮肤分泌物进行一系列层析分离，最终纯化 得到4种抗菌肽，分别命名为Odorgrin A、Odorgrin B、Odorgrin C和Odorgrin D．质谱测得单一同位素分子量 分别为 3 364．8、3 817．9、4 859．4和3 287．9．Edman降解法测得OdorgrinA的氨基酸序列为GLLDTFKNLALNAAKSAGVSV— LNSLSCKLSKTC．将以上结果与国内外对该物种抗菌肽研究的已有结果进行了比较分析，证明本研究分离到的4种抗 菌肽与前人分离到的部分抗菌肽间具有一致性．图3表3参25 关键词 无指盘臭蛙；皮肤分泌物；抗菌肽；分离纯化；氨基酸序列 CLC Q959．530．6 Purification and Amino Acid Sequencing of Antimicrobial Peptides from Odorrana grahami Skin LIU Jiongyu，JIANG Jianping，HE Kaize，XIE FengZHENG Zhonghua (ChengduInstitute ofBiology，ChineseAcademy ofSciences，Chengdu 610041，China) Abstract The skin secretion was produced by electrical shock stimulating the body of the diskless—fingered odorous frog (Odorrana grahami)caught from Mianning County，Sichuan Province．Four antimicrobial peptides，named Odorgrin A，Odor— grin B，Odorgrin C and Odorgrin D，were separated and purified from the secretion by CM—sepharose fast flow ion exchange chromatography，sephadexG-25 exclusion chromatography，SOUCE 15 RPC reverse phase chromatography and C18 reverse phase HPLC successively．The monoisotopic molecular masses of Odorgrin A，Odorgrin B，Odorgrin C and Odorgrin D were measured by ESI—MS as 3 364 ．8，3 817．9，4 859．4 and 3 287．9，respectively．The amino acid sequence of Odorgrin A was G DTFKNLALNAAKSAGVSVLNSLSCKISKTC obtained by automated Edman degradation sequencing．The properties of these 4 antimicrobial peptides were compared with those obtained from the same species by other researchers．Fig 3，Tab 3， Ref 25 Keywords Odorrana grahami；sk