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Peptides Biology and Chemistry

:	Xiao-Yu Hu, Rui Wang, James P. Tam
:	Peptides Biology and Chemistry
:	Kluwer Academic Publishers
:	9780306468803
:	1
:	CHF 109.20
:

:	Naturwissenschaft
:	English

:	290
:	DRM
:	PC/MAC/eReader/Tablet
:	PDF

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.

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.

	Preface	5
	Contents	11
	Abbreviations	19
	Session A Novel approach on synthetic method	21
	Orthogonal ligation of free peptides	22
	Solid phase synthesis of peptide aldehydes	27
	Investigation of enzyme activity and inhibition in the interior of novel solid supports	33
	Immobilization of a-Chymotrypsin on zeolite for peptide synthesis in organic solvent	40
	Preparation of a peptide thioester using a fluoren-9-ylmethoxycarbonyl solid-phase method	45
	Application of DEPBT for the synthesis of N-protected peptide alcohols	49
	Decomposition of amino acid cupric complex by using tetrahydrothiazole-2-thione for preparation ofN a-Boc-Ne-Fmoc-L-Lysine	52
	Two-step selective formation of three disulfide bridges in the synthesis of d-Gonotoxin PVIA	55
	Session B Structure, folding and conformation analysis De novo design of peptide	55
	59	55
	Design, synthesis and NMR structure of a rigid cyclic peptide template	60
	Automated docking of Sch68631 with HCV NS3 protease	65
	Molecular modeling of the three-dimensional structure of the human interleukin-11	68
	Designing ß-hairpin forming short peptide	72
	Anti-tumor activities of papavor somniferum pollen peptides	76
	Structural investigation of two pollen peptides by 2 D homonuclear NMR spectroscopy	79
	A comparison of three heuristic algorithms for molecular docking	82
	Aggregation of a calmodulin-binding peptide	86
	Session C Neuro/Endocrino/Bioactive peptide	90
	Neuropeptide AVP(4-8) playing an indirect neurotrophic role	91
	Potent bradykinin antagonists having medical potential	93
	Understanding the chemistry and biology of rodent relaxin: Role of the C-peptide	97
	Purification and characterization of a new conotoxin from the venom ofConus betulinus	102
	Synthesis and biological activity of human calcitonin analogs	105
	Enhanced expression of CNDF mRNA in rat brain by administration of AVP(4-8)	108
	The synthesis and opioid activities of nociceptin and its fragments	111
	The effect of Glu position change on the opioid activity of deltorphin II	114
	Isolation and purification of a series of peptides from Panax notoginseng	118
	Study of the relationship between structure and function of HIV-1 gp 41 N terminus fusion peptide	120
	Hypotensive activities of nociceptin and its fragments	124
	The design and synthesis of endomorphins and their analogues	128
	Neuronal apoptosis induced by ß-amyloid peptides in vitro	132
	Studies on the spin-labeling technique on deltorphin II and its analogues	135
	Design and synthesis of salmon calcitonin and its analogues	140
	Studies on the synthesis and antithrombosis activity of two peptides derived from the molecular-binding site of fibrinogen	142
	Synthesis and expression of w-conotoxin MVIIA	145
	Session D Peptide vaccines/Immunology/Virus	150
	Studies on the synthetic peptide vaccines against schistosomiasis	151
	Studies on the anti-schistosomal synthetic peptide vaccine: Prediction and synthesis of antigenic peptides of Schistosoma paramyosin	154
	Construction and expression of a recombinant antibody targeted plasminogen activator	156
	Chemical conjugation between haemophilus influenzae type b (hib) polysaccharide and proteins	159
	Synthesis of human inhi