Hu / Wang / Tam Peptides

Biology and Chemistry
1. Auflage 2006
ISBN: 978-0-306-46880-3
Verlag: Springer Netherland
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 290 Seiten, eBook

Reihe: Chinese Peptide Symposia

The Fifth Chinese Peptide Symposium, hosted by Lanzhou University, was held at Lanzhou, China July 14-17, 1998, with 156 participants, including 30 scientists from abroad, representing nine countries. The four-day conference was both intense and spiritually rewarding. Our goal for CPS-98 was to provide a forum for the exchange of knowledge, cooperation and friendship between the international and Chinese scientific communities, and we believe this goal was met. The symposium consisted of 8 sessions with 42 oral and 90 poster presentations, including synthetic methods, molecular diversity and peptide libraries, structure and conformation of peptides and proteins, bioactive peptides, peptide immunology, De Novo design and synthesis of proteins and peptides, ligand-receptor interactions, the chemistry-biology-interface and challenging problems in peptides. The enthusiastic cooperation and excellent contributions were gratifying and the active response of the invited speakers contributed to the success of the symposium. The presentations were of excellent caliber and represented the most current and significant aspects of peptide science. Dr. Kit Lam of the University of Arizona and Dr. Yun-Hua Ye of Peking University were the recipients of “The Cathay Award” sponsored by the H. H. Liu Education Foundation, offered for their seminal contribution in peptide science and the Chinese Peptide Symposium. Four outstanding young scientists were selected by the organizing committee to receive awards sponsored by Haikou Nanhai Pharmaceutical Industry Co. Ltd. (Zhong He Group).

Zielgruppe

Research

Autoren/Hrsg.

Hu, Xiao-Yu

Wang, Rui

Tam, James P.

Inhaltsverzeichnis

Preface. Chinese Peptide Symposium - 1998. Session A: Novel approach on synthetic method. Orthogonal ligation of free peptides; J.P. Tam, et al. Solid phase synthesis of peptide aldehydes; J.-A. Fehrentz, et al. Investigation of enzyme activity and inhibition in the interior of novel solid supports; M. Meldal, et al. Immobilization of alpha-chymotrypsin on zeolite for peptide synthesis in organic solvent; Y.-H. Ye, et al. Preparation of a peptide thioester using a fluoren-9-ylmethoxycarbonyl solid-phase method; X.-Q. Li. Application of DEPBT for the synthesis of N-protected peptide alcohols; G.-L. Tian, et al. Decomposition of amino acid cupric complex by using tetrahydrothiazole-2-thione for preparation of Nalpha-Boc-Nepsilon-Fmoc-L-Lysine; X.-M. Gao, et al. Two-step selective formation of three disulfide bridges in the synthesis of delta-Conotoxin P VIA; H. Jiang, et al. Session B: Structure, folding and conformation analysis/De novo design of peptide & protein. Design, synthesis and NMR structure of a rigid cyclic peptide template; S.A. Muhle, J.P. Tam. Automated docking of Sch68631 with HCV NS3 protease; X.-J. Xu. Molecular modeling of the three-dimensional structure of the human interleukin-11; L.-R. Chen, et al. Designing beta-hairpin forming short peptide; Y.-L. Sha, et al. Anti-tumor activities of papavor somniferum pollen peptides; J.-X. Xu, S. Jin. Structural investigation of two pollen peptides by 2D homonuclear NMR spectroscopy; Y.-L. Song, et al. A comparison of three heuristic algorithms for molecular docking; T.-J. Hou, et al. Aggregation of a calmodulin-binding peptide; H.-S. Yan, et al. Session C: Neuro/Endocrino/Bioactive peptide. Neuropeptide AVP(4-8) playing an indirect neurotrophic role; Q.-W. Yan, et al. Potent bradykinin antagonists having medical potential; J.M. Stewart, et al. Understanding the chemistry and biology of rodent relaxin: Role of the C-peptide; J.D. Wade, et al. Purification and characterization of a new conotoxin from the venom of Conus betulinu; C.-X. Fan, et al. Synthesis and biological activity of human calcitonin analogs; C. Yu, et al. Enhanced expression of CNDF mRNA in rat brain by administration of AVP(4-8); W.-X. Li, et al. The synthesis and opioid activities of nociceptin and its fragments; S.-L. Dong, et al. The effect of Glu position change on the opioid activity of deltorphin II; R. Wang, et al. Isolation and purification of a series of peptides from Panax notoginseng; J.-G. Ji, et al. Study of the relationship between structure and function of HIV-1 gp 41 N terminus fusion peptide; M. Wu, et al. Hypotensive activities of nociceptin and its fragments; R. Wang, et al. The design and synthesis of endomorphins and their analogues; X.-F. Huo, et al. Neuronal apoptosis induced by &bgr;-amyloid peptides in vitro; W.-X. Li, et al. Studies on the spin-labeling technique on deltorphin II and its analogues; D.-J. Yang, et al. Design and synthesis of salmon calcitonin and its analogues; H.-P. Pan, et al. Studies on the synthesis and antithrombosis activity of two peptides derived from the molecular--binding site of fibrinogen; X.-Y. Hu. Synthesis and expression of &ohgr;-conotoxin MVIIA; M.-Y. Cai, X.-M. Qu. Session D: Peptide Vaccines/Immunology/Virus. Studies on the synthetic peptide vaccines against schistosomiasis; S.-L. Cao, et a

Leseproben

Detection of gene expression product of transgenic tobacco by antigenic peptide (p. 215-216)

Jia-Xi Xu a*, Yuan Ma b, Mi Ma c and Zhong-Ping Lin d
aDepartment of Chemistry, Peking University. Beijing, 100871,
bDepartment of Chemistry, Tsinghua University, Beijing, 100084,
cInstitute of Botany, Academia Sinica, Beijing 10044,
dDepartment of Biology, Peking University, Beijing, 100871, China

Introduction

The need as well as the ability to cultivate special plants, such as antiviral wheat, antiinsect plants, economical plants etc, has increased with advances in biological science and technology and has created significant economic benefits. It is easy to detect DNA and RNA in transgenic processing. However, it is difficult to detect the protein as gene expression product due to low concentration and difficult separation and purification (1). Now we have used a synthetic antigenic peptide to solve this problem. After predicting and synthesizing an antigenic peptide of isopentyl transferase in transgenic tobacco, a key enzyme in gene expression, a peptide with the sequence IHARQQEQKF was conjugated to BSA. Its antibodies were then obtained from rabbit sera after immunizing the rabbit with this conjugated antigen. The antibody can be used for the qualitative and quantitative determination of gene expression product in crude proteinextract of leaf, stem and root of transgenic tobacco by ELISA and Western blot methods.

Results and Discussion
Prediction of epitope

The nucleotide and amino acid sequences of the isopentyl transferase in transgenic tobacco was obtained from published data (2). Its epitope was predicted according to the hydrophilicity (Hopp and Woods method), flexibility and accessibility by the computer program PC-Gene (3,4). Its secondary structure was predicted by the Chou and Fasman method (5). Peptide IHARQQEQKF (212-221 amino acid residues) with high hydrophilicity, flexibility and accessibility was predicted.

Synthesis of epitopic peptides

The predicted peptide was synthesized by Merrifield solid phase peptide synthesis method with the acid-labile tert-butyloxycarbonyl (Boc) group for temporary protection and acid-stable groups for side chain protection (3, 4, 6). Side chain-protected peptideresin was cleaved by anhydrous hydrogen fluoride under anisole, 1, 2-ethandithiol and thioanisole as scavengers and washed with cooled ethyl ether. Peptide was extracted with 30% acetic acid and purified by gel filtration on Sephadex G10. It was further purified by preparative HPLC and checked for homogeneity by reverse-phase HPLC. Its structure was confirmed by amino acid analysis and FAB-MS.

Immunologiccal methods

This antigenic peptide was conjugated to BSA by the glutaraldehyde method and its antibodies were then obtained from rabbit sera after immunizing rabbit with this conjugated antigen (7). The antibody was used to detect the gene expression product of transgenic tobaco at a dilution of 1:132 with OD492 nm 0.95 by the ELISA method. The results are shown in Table 1. This is a simple method for the qualitative and quantitative determination of gene expression product in crude protein-extract of leaf, stem and root of transgenic tobacco using ELISA and Western blot methods.