EPITOPE MAPPING AND ANALYSIS OF A GROWTH-ENHANCING MONOCLONAL-ANTIBODY BY LIMITED TRYPTIC DIGESTION OF PORCINE GH

被引:10
作者
SHIEH, HM
BASS, RT
WANG, BS
CORBETT, MJ
BUCKWALTER, BL
机构
[1] Agricultural Research Division, American Cyanamid Company, Princeton
关键词
D O I
10.1677/joe.0.1450169
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
In this study, the epitope of a murine PS-7.6 monoclonal antibody (mAb) which was raised against the recombinant porcine GH (pGH) and subsequently shown to enhance the growth-promoting activity of pGH in a hypophysectomized rat model, was mapped by the limited tryptic digestion of pGH. A pGH fragment corresponding to amino acid residues 70-95 was separated by reverse-phase HPLC and also immunoprecipitated by PS-7.6 mAb. This fragment was found in an RIA to compete with radiolabelled pGH for the binding of PS-76 mAb in a dose-dependent fashion. Several peptides covering this potential epitope region of pGH(70-95) were synthesized and assayed by competitive RIA. The results suggested that pGH(75-90) was the optimal sequence recognized by PS-7.6 mAb. Sequential alanine substitution of each residue of pGH(75-90) revealed that the side chains of Leu(76) Ile(83) and Leu(87) were critical for binding to PS-7.6 mAb. Other residues could be replaced by alanine without substantially altering the binding affinity. The region of amino acids 75-95 comprises the C-terminal end of the second helix of pGH and the repeating pattern of i and i+3 (i+7) of the critical amino acids appears consistent with PS-76 mAb binding to the hydrophobic side of the helix. The sequence and the helical structure of the epitope of PS-76 mAb provide the basis for designing the effective peptide vaccines to enhance the growth performance of animals.
引用
收藏
页码:169 / 174
页数:6
相关论文
共 29 条
[1]   3-DIMENSIONAL STRUCTURE OF A GENETICALLY ENGINEERED VARIANT OF PORCINE GROWTH-HORMONE [J].
ABDELMEGUID, SS ;
SHIEH, HS ;
SMITH, WW ;
DAYRINGER, HE ;
VIOLAND, BN ;
BENTLE, LA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (18) :6434-6437
[2]   POTENTIATION OF THE SOMATOGENIC AND LACTOGENIC ACTIVITY OF HUMAN GROWTH-HORMONE WITH MONOCLONAL-ANTIBODIES [J].
ASTON, R ;
HOLDER, AT ;
PREECE, MA ;
IVANYI, J .
JOURNAL OF ENDOCRINOLOGY, 1986, 110 (03) :381-388
[3]   ENHANCEMENT OF BOVINE GROWTH-HORMONE ACTIVITY INVIVO BY MONOCLONAL-ANTIBODIES [J].
ASTON, R ;
HOLDER, AT ;
IVANYI, J ;
BOMFORD, R .
MOLECULAR IMMUNOLOGY, 1987, 24 (02) :143-150
[4]   TERMINATION OF IMMUNOLOGICAL UNRESPONSIVENESS TO BOVINE SERUM ALBUMIN IN RABBITS .3. STRUCTURAL AND SEROLOGICAL RELATIONSHIPS AMONG VARIOUS SERUM ALBUMINS AND THEIR CYANOGEN BROMIDE FRAGMENTS [J].
BENJAMIN, DC ;
WEIGLE, WO .
IMMUNOCHEMISTRY, 1971, 8 (12) :1087-&
[5]  
BURT SK, 1988, ANNU REP MED CHEM, V23, P285
[6]  
CAMPBELL RG, 1991, J ANIM SCI, V69, P1522
[7]   EMPIRICAL PREDICTIONS OF PROTEIN CONFORMATION [J].
CHOU, PY ;
FASMAN, GD .
ANNUAL REVIEW OF BIOCHEMISTRY, 1978, 47 :251-276
[8]   IMMUNOLOGICAL ACTIVITY OF SOME OF CHYMOTRYPTIC PEPTIDES OF SPERM-WHALE MYOGLOBIN [J].
CRUMPTON, MJ ;
WILKINSO.JM .
BIOCHEMICAL JOURNAL, 1965, 94 (03) :545-&
[9]   HIGH-RESOLUTION EPITOPE MAPPING OF HGH-RECEPTOR INTERACTIONS BY ALANINE-SCANNING MUTAGENESIS [J].
CUNNINGHAM, BC ;
WELLS, JA .
SCIENCE, 1989, 244 (4908) :1081-1085
[10]   DOMINANT FORCES IN PROTEIN FOLDING [J].
DILL, KA .
BIOCHEMISTRY, 1990, 29 (31) :7133-7155