Hu / Wang / Tam Peptides

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

E-Book, Englisch, 290 Seiten

Reihe: Chinese Peptide Symposia

This volume summarizes the papers presented at the Chinese Peptide Symposium-96. The topics of the Symposium comprised six sections:

- approach and methods,
- a molecular diversity-peptide library,
- conformation of peptides and proteins,
- structure and design,
- bioactive peptides, and
- a chemistry-biology interface.

The contributions were concerned with most aspects of peptides and proteins, including chemistry and biology, along with others related to peptides, such as glycopeptides, peptidomimetics and amino acids. The proceedings reflect recent advances in the fields of peptide and protein chemistry and biology, providing a stimulus for continuing research in these and related fields.

Autoren/Hrsg.

Hu, Xiao-Yu

Wang, Rui

Tam, James P.

Inhaltsverzeichnis

1;Preface;5
2;Contents;11
3;Abbreviations;19
4;Session A Novel approach on synthetic method;21
4.1;Orthogonal ligation of free peptides;22
4.2;Solid phase synthesis of peptide aldehydes;27
4.3;Investigation of enzyme activity and inhibition in the interior of novel solid supports;33
4.4;Immobilization of a-Chymotrypsin on zeolite for peptide synthesis in organic solvent;40
4.5;Preparation of a peptide thioester using a fluoren-9-ylmethoxycarbonyl solid-phase method;45
4.6;Application of DEPBT for the synthesis of N-protected peptide alcohols;49
4.7;Decomposition of amino acid cupric complex by using tetrahydrothiazole-2-thione for preparation ofN a-Boc-Ne-Fmoc-L-Lysine;52
4.8;Two-step selective formation of three disulfide bridges in the synthesis of d-Gonotoxin PVIA;55
5;Session B Structure, folding and conformation analysis De novo design of peptide & protein;59
5.1;Design, synthesis and NMR structure of a rigid cyclic peptide template;60
5.2;Automated docking of Sch68631 with HCV NS3 protease;65
5.3;Molecular modeling of the three-dimensional structure of the human interleukin-11;68
5.4;Designing ß-hairpin forming short peptide;72
5.5;Anti-tumor activities of papavor somniferum pollen peptides;76
5.6;Structural investigation of two pollen peptides by 2 D homonuclear NMR spectroscopy;79
5.7;A comparison of three heuristic algorithms for molecular docking;82
5.8;Aggregation of a calmodulin-binding peptide;86
6;Session C Neuro/Endocrino/Bioactive peptide;90
6.1;Neuropeptide AVP(4-8) playing an indirect neurotrophic role;91
6.2;Potent bradykinin antagonists having medical potential;93
6.3;Understanding the chemistry and biology of rodent relaxin: Role of the C-peptide;97
6.4;Purification and characterization of a new conotoxin from the venom ofConus betulinus;102
6.5;Synthesis and biological activity of human calcitonin analogs;105
6.6;Enhanced expression of CNDF mRNA in rat brain by administration of AVP(4-8);108
6.7;The synthesis and opioid activities of nociceptin and its fragments;111
6.8;The effect of Glu position change on the opioid activity of deltorphin II;114
6.9;Isolation and purification of a series of peptides from Panax notoginseng;118
6.10;Study of the relationship between structure and function of HIV-1 gp 41 N terminus fusion peptide;120
6.11;Hypotensive activities of nociceptin and its fragments;124
6.12;The design and synthesis of endomorphins and their analogues;128
6.13;Neuronal apoptosis induced by ß-amyloid peptides in vitro;132
6.14;Studies on the spin-labeling technique on deltorphin II and its analogues;135
6.15;Design and synthesis of salmon calcitonin and its analogues;140
6.16;Studies on the synthesis and antithrombosis activity of two peptides derived from the molecular-binding site of fibrinogen;142
6.17;Synthesis and expression of w-conotoxin MVIIA;145
7;Session D Peptide vaccines/Immunology/Virus;150
7.1;Studies on the synthetic peptide vaccines against schistosomiasis;151
7.2;Studies on the anti-schistosomal synthetic peptide vaccine: Prediction and synthesis of antigenic peptides of Schistosoma paramyosin;154
7.3;Construction and expression of a recombinant antibody targeted plasminogen activator;156
7.4;Chemical conjugation between haemophilus influenzae type b (hib) polysaccharide and proteins;159
7.5;Synthesis of human inhibin fragments, preparation and generation of monoclonal antibodies against human inhibin;161
8;Session E Peptide diversity; Chemical libraries; Ligand-receptor interaction; Peptides in therapeutics & diagnosis;165
8.1;Design of a d opioid peptide mimetic based on a topographically constrained cyclic enkephalin;166
8.2;Tetrazole cis-amide bond mimetics identify the ß-turn conformation of insect kinin neuropeptides;171
8.3;Synthetic studies of farnesyl protein transferase inhibitor pepticinnamin E;176
8.4;Novel coupling reagents;179
8.5;Pharmacophore model of growth hormone secretagogues;182
8.6;Discovery that deltorphin II derivatives are potent melanotropins, putatively active at the Xenopus melanocortin- 1 receptor;185
9;Session F Glyco/Lipo/Phospho peptide; Peptide diversity/Chemical libraries; Receptor-ligand interaction;188
9.1;Enzymatic synthesis of phosphopeptides;189
9.2;Synthesis of phosphorylated polypeptide by a thioester method;192
9.3;Difference between N-phospho-L- and D-methionine in ester exchange reaction with uridine;198
9.4;Synthesis of analogs of phosphoamino acids and their biomimic reactions;200
9.5;Solid phase synthesis of heterocyclic combinatorial libraries derived from peptides;204
9.6;Parallel synthesis of muramyl peptides derivatives;210
9.7;ß-Cyclodextrin for presentation of bioactive peptides to molecular recognition;214
9.8;The C-terminus of d-opioid receptor does not control DPDPEinduced desensitization;222
10;Session G Peptides in therapeutics and diagnosis;225
10.1;Detection of gene expression product of transgenic tobacco by antigenic peptide;226
10.2;Advantages of Calcitonin complexed with SA liposome;228
10.3;Hypoglycemic effects of insulin sublingual drops containing azone on rats and rabbits;232
11;Session H Other research related to peptides and proteins;235
11.1;Secretory expression of porcine insulin precursor in methylotrophic yeast Pichia pastoris;236
11.2;Chemo-enzymatic syntheses of eel calcitonin analogs having natural N-linked oligosaccharides;239
11.3;Design of specific inhibitors at the ATP binding site of DNA topoisomerases II;243
11.4;Applications of optical biosensors to structure-function studies in the EGF/EGF receptor system;247
11.5;Experimental study of osteogenic growth peptide promoting bone mass in rat osteoporosis;253
11.6;Determination of inhibitory constants for CPA by competitive spectrophotometry;256
11.7;Sublingual administration of monomeric insulin - destetrapeptide insulin;258
11.8;How to get patent rights to protect your invention in biotechnology;261
11.9;Study on peptide maps from milk protein hydrolysates by SEHPLC;264
11.10;Synthesis and solution structures of new chiral enantiomeric peptide nucleic acid (PNA) dimers;267
11.11;Study of one type of metallothionein and SZ51 recombinant protein directing and imaging reagent;271
11.12;LHRH antagonists: new preclinical and clinical results;273
11.13;DNA-binding characteristics of antitumor drug actinomycin D and its analogs;278
11.14;Design, synthesis of the peptide analogs of antitumor agent actinomycin D;283
12;Authors Index;288
13;Subject Index;291

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.