title: The role of the N-terminal leucine residue in snake venom cardiotoxin II (Naja naja atra).
abstract: The N-terminal leucine residue of snake venom cardiotoxin II (CTX II) (Naja naja atra) was systematically replaced with D-leucine (CTXII-L1-D-L), glycine (CTXII-L1G) or deleted [CTXII-(2-60)] to study the role of leucine residue in CTX II molecule. CTX II, CTXL1-D-L, CTXL1G and CTX(2-60) were produced by chemical synthesis method and purified by high performance liquid chromatography. Owing to folding problem in CTXII-(2-60), only CTX II, CTXII-L1-D-L and CTXII-L1G were produced in a pure form and characterized by amino acid analysis, mass spectrometry and peptide mapping. In the structural aspect, changing the Leu-1 by D-Leu or Gly causes a drastic alteration in the whole CTX II structure as detected by circular dichroism, 1-anilino-naphthalene-8-sulfonate (ANS) fluorescence assay. In the functional aspect, both CTXII-L1-D-L and CTXII-L1G are still retained substantial biological activity of CTX II. Therefore, the results indicate that both the chirality and the side-chain of the N-terminal leucine residue of CTX II are important elements in maintaining the whole CTX II structure. In addition, this study is the first report in elucidating the reason why the first N-terminal residue of most CTXs (90.3%) is leucine residue.
authors.affiliation: Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, Republic of China.
authors.forename: C Y
authors.initials: CY
authors.lastname: Wu
authors.forename: W C
authors.initials: WC
authors.lastname: Chen
authors.forename: C L
authors.initials: CL
authors.lastname: Ho
authors.forename: S T
authors.initials: ST
authors.lastname: Chen
authors.forename: K T
authors.initials: KT
authors.lastname: Wang
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002942:Circular Dichroism; D004179:Cobra Cardiotoxin Proteins / Q000737:chemistry* / Q000633:toxicity; D007930:Leucine / Q000737:chemistry; D051379:Mice; D015394:Molecular Structure; D010449:Peptide Mapping; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D000596:Amino Acids; D004179:Cobra Cardiotoxin Proteins; D007930:Leucine