Enzymic and immunochemical properties of lysozyme XVI. A novel synthetic approach to an antigenic reactive site by direct linkage of the relevant conformationally adjacent residues constituting the site

Atassi, M. Zouhair; Lee, Ching-Li; Pai, Rong-Chang

doi:10.1016/0005-2795(76)90219-1

Cited by 65 publications

(29 citation statements)

References 24 publications

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“…So we examined the covalent structure of bovine serum albumin (Brown, 1975), bearing in mind that functional equivalence between two antigenic sites implies resemblance in binding capacity and would not necessarily require complete structural identity (Atassi et al, 1976b;Lee et al, 1976). We concluded that the regions 137-146, 328-337 and 525-534 (Fig.…”

Section: Discussionmentioning

confidence: 99%

Localization and verification by synthesis of five antigenic sites of bovine serum albumin

Atassi¹,

Sakata²,

Kazim³

1979

Biochemical Journal

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Recently we have shown that the major antigenic sites of bovine serum albumin exhibit functional equivalence progressively increasing with the time at which antibodies are obtained after the first immunization. Analysis of our recent immunochemical findings and the known covalent structure of bovine serum albumin have enabled us to predict the locations of five antigenic sites of bovine serum albumin. The predicted locations were synthesized, and immunochemical studies with late-course antisera showed them to constitute antigenic sites of native bovine serum albumin.

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Section: Discussionmentioning

confidence: 99%

Localization and verification by synthesis of five antigenic sites of bovine serum albumin

Atassi¹,

Sakata²,

Kazim³

1979

Biochemical Journal

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“…Under the conditions described above, the rate of hydrolysis of TAME by the final TrPepz design (Fig. 4) was better than designs [1][2][3] by 250, 123, and 72 times, respectively. The peptide maps of 144-hr Mb or casein hydrolysates by designs 2 and 3 coma 10.27 13.16 -!,~19…”

Section: Discussionmentioning

confidence: 99%

“…X-ray crystallography of enzymes and their complexes with substrate analogs or inhibitors has provided detailed information about the architecture and contact residues ofthe active sites of many enzymes. In 1976, Atassi and coworkers (1,2) devised the technique of "surfacesimulation" synthesis, in which the spatially adjacent residues constituting a protein binding site are directly linked via peptide bonds with appropriate spacing and directionality. A peptide is thus generated that does not exist in the protein but mimics the conformation and disposition of the residues of the binding site.…”

mentioning

confidence: 99%

Design of peptide enzymes (pepzymes): surface-simulation synthetic peptides that mimic the chymotrypsin and trypsin active sites exhibit the activity and specificity of the respective enzyme.

Atassi

Manshouri

1993

Proc. Natl. Acad. Sci. U.S.A.

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Two 29-residue peptides were prepared, one of which (ChPepz) was designed by surface-simulation synthesis to mimic the active site of a-chymotrypsin, and the other (TrPepz), which contained four substitutions relative to ChPepz, was fashioned after the active site of trypsin. Each peptide was cyclized by a disulfide bond. The ChPepz monomer effected hydrolysis of the ester group in N-benzoyl-L-tyrosine ethyl ester, an a-chymotrypsin substrate, with Km and kt values that were comparable to those of a-chymotrypsin. ChPepz was completely inactivated by diisopropyl fluorophosphate (DIFP), L-1-p-tosylamino-2-phenylethyl chloromethyl ketone (TPCK), or reduction of the disulfide bond. It had no catalytic activity on N-tosyl-L-arginine methyl ester, a trypsin substrate. On the other hand, TrPepz, which had no effect on N-benzoyl-L-tyrosine ethyl ester, hydrolyzed N-tosyl-L-arginine methyl ester with a Km value that was essentially identical to that of trypsin, but its k5,,, value was almost half that of trypsin. TrPepz was fully inactivated by reduction of the disulfide bond, by DIFP, or by phenylmethylsulfonyl fluoride but not by TPCK. It was also completely inhibited by soybean trypsin inhibitor, bovine pancreatic trypsin inhibitor, and human a1-antitrypsin. ChPepz and TrPepz hydrolyzed proteins (myoglobin and casein) to give panels ofpeptides that were similar to those of the same protein obtained with the respective enzyme. However, TrPepz was more efficient than trypsin at hydrolyzing the C bonds of two or more consecutive lysine and/or arginine residues. Like its esterase activity, the proteolytic activity of ChPepz was inhibited by either DIFP or TPCK whereas that of TrPepz was inhibited by either DIFP or phenylmethylsulfonyl fluoride but not by TPCK. Finally, ChPepz and TrPepz were each more active at low temperature than the respective enzyme. This ability to construct fully functional peptide enzymes (pepzymes) of chosen specificities should find many practical applications.The critical role of enzymes in the catalysis of biological processes has, for decades, made enzymes the subject of the most intense studies. The duplication of the activities of enzymes by synthetic analogs has been a prime goal of biochemists. X-ray crystallography of enzymes and their complexes with substrate analogs or inhibitors has provided detailed information about the architecture and contact residues ofthe active sites of many enzymes. In 1976, Atassi and coworkers (1, 2) devised the technique of "surfacesimulation" synthesis, in which the spatially adjacent residues constituting a protein binding site are directly linked via peptide bonds with appropriate spacing and directionality. A peptide is thus generated that does not exist in the protein but mimics the conformation and disposition of the residues of the binding site. Surface-simulation synthesis (for review, see ref.3) has been employed to mimic protein antigenic sites (1, The publication costs of this article were defrayed in part by page charge payment. This arti...

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“…The discontinuous nature of the lysozyme antigenic sites (see Ref. [3] for definition of term) was confirmed by surfacesimulation synthesis [4,5]. In this approach, the functional sites, which were composed of spatially-close residues that may not be directly linked by peptide bonds in the sequence, could be mimicked by a synthetic peptide that contained the active site residues with suitable spacing and directionality.…”

Section: Introductionmentioning

confidence: 99%

Antibodies against a synthetic peptide designed to mimic a surface area of the H chain of botulinum neurotoxin A

et al. 2012

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Enzymic and immunochemical properties of lysozyme XVI. A novel synthetic approach to an antigenic reactive site by direct linkage of the relevant conformationally adjacent residues constituting the site

Cited by 65 publications

References 24 publications

Localization and verification by synthesis of five antigenic sites of bovine serum albumin

Localization and verification by synthesis of five antigenic sites of bovine serum albumin

Design of peptide enzymes (pepzymes): surface-simulation synthetic peptides that mimic the chymotrypsin and trypsin active sites exhibit the activity and specificity of the respective enzyme.

Antibodies against a synthetic peptide designed to mimic a surface area of the H chain of botulinum neurotoxin A

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