ImmunoMethods最新文献

{"title":"Identification of Interactive Sites of Proteins and Protein Receptors by Computer-Assisted Searches for Complementary Peptide Sequences","authors":"Fassina Giorgio,&nbsp;Melli Marialuisa","doi":"10.1006/immu.1994.1045","DOIUrl":"10.1006/immu.1994.1045","url":null,"abstract":"<div><p>Protein sites important for receptor binding have been identified in several systems by searching for protein/receptor stretches characterized by hydropathic anti-complementarity. A computer-assisted method [SITESEARCH] has been developed to identify protein sites responsible for receptor recognition, once the amino acid sequences of the protein ligand and its receptor are available. The software first determines the hydropathic profiles of the two polypeptide chains under investigation, and then compares profiles of preselected length, determining at the same time the degree of their hydropathic complementarity. The procedure is repeated until all the different segments in the two chains are compared. Fragments characterized by the maximal level of hydropathic complementarity are selected as putative binding sites. This approach has been initially applied to the interleukin-1β (IL-1β)/receptor type I case. SITESEARCH identified residues 88-99 in IL-1β and 151-162 in the receptor as the sequence pair characterized by the maximal level of hydropathic complementarity. These fragments, once produced by chemical synthesis, have displayed specific recognition properties for each other, as detected by solid-phase binding assays. The IL-1β sequence identified corresponds to a highly exposed part of the protein molecule, and substitution of this sequence with another of the same length but with different hydropathic characteristics generated mutants with drastically reduced binding activity to the receptor. Mutations in this sequence did not alter the protein biological activity, thus suggesting the structural integrity of the mutants. Cumulatively, these results validate the SITESEARCH prediction, suggesting that IL-1β sequence 88-99 is involved in at least a portion of the protein/receptor binding site.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 114-120"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1045","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of Complementary Peptides and Their Antibodies in T-Cell-Mediated Autoimmune Disease: Experiments with Myelin Basic Protein","authors":"Zhou Shan-Ren,&nbsp;Whitaker John N.","doi":"10.1006/immu.1994.1048","DOIUrl":"10.1006/immu.1994.1048","url":null,"abstract":"<div><p>In this article we review the field of idiotype (Id) network and experimental allergic encephalomyelitis, focusing on the generation of monoclonal antibody anti-Id by using complementary peptide and the investigation of anti-Id immunoregulatory effects on immune responses of B and T cells to specific myelin basic protein peptides <em>in vitro</em> and <em>in vivo</em>.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 136-147"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18541906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Complementary Peptides as Probes to Explore Neuropeptide Receptors on Lymphocytes","authors":"Johnson Howard M.,&nbsp;Torres Barbara A.","doi":"10.1006/immu.1994.1051","DOIUrl":"10.1006/immu.1994.1051","url":null,"abstract":"<div><p>Studies on neuroendocrine hormone receptor have been hampered by low numbers and concentrations of receptors found within and outside the neuroendocrine system. The complementary peptide approach is particularly useful for dealing with this problem and has been used to characterize lymphoid receptors for arginine vasopressin (AVP), corticotropin (ACTH), substance P, and opioid peptides. A nonapeptide derived by reading of the complementary DNA strand of the bovine AVP gene in the 3′ to 5′ direction specifically blocks the AVP helper signal for interferon-γ production by mouse T lymphocytes. Antibodies to 3′-5′ AVP-binding peptide bound to cells, and the binding was inhibited by excess AVP. Thus, binding of anti-3′-5′ AVP-binding peptide antibodies to the AVP receptor was specific. The complementary peptide approach has also been used to produce antibodies specific for the ACTH receptor complex. Complementary peptides to ACTH derived by reading in either the 5′ to 3′ or 3′ to 5′ direction were able to bind to ACTH. Mono-specific antibodies to the ACTH(1-24) complementary peptide caused an ACTH-like steroidogenic response of cultured mouse adrenal cells, presumably by binding to the ACTH receptor, and binding was specifically inhibited by ACTH. The ACTH receptor complex from solubilized adrenal cells was shown to consist of four subunits with <em>M</em>_r 83,000, 64,000, 52,000, and 22,000. The 83,000 and 52,000 <em>M</em>_r subunits are disulfide linked and noncovalently associated with the other subunits, with binding of labeled ACTH localized to the 83,000 <em>M</em>_r subunit. Similarly, a complementary peptide was shown to bind directly to substance P in a saturable and dose-dependent manner. Affinity-purified antibodies to the substance P complementary peptide directly bound to IM-9 cells and blocked the binding of substance P to its IM-9 receptor. Further, these antibodies recognized a single protein of 58 kDa from solubilized IM-9 cells. Complementary peptides have been used to characterize opioid receptors, and studies show that antigenic and functional similarity exists between opioid receptors from brain and lymphocytes. Thus, the complementary peptide approach can be used to produce antibodies specific for neuroendocrine receptors, which are useful tools for the isolation and characterization of these receptors.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 167-171"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1051","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Complementary Peptides as Antibody Mimetics for Protein Purification and Assay","authors":"Fassina Giorgio","doi":"10.1006/immu.1994.1046","DOIUrl":"10.1006/immu.1994.1046","url":null,"abstract":"<div><p>The possibility of designing sequence-directed recognition peptides (complementary peptides) able to noncovalently associate target peptides or proteins is one of the most important biotechnological applications deriving from the molecular recognition theory [MRT]. Complementary peptides can be used widely as synthetic ligands for the development of affinity purification strategies to isolate target peptides or proteins from crude sources. Generally, the observed affinity and selectivity are sufficient to allow one-step purification of target molecules directly from crude mixtures and the possibility of producing the ligands in enzymatically stable forms greatly enhance their applicability. In addition, the noninterference of nonionic detergents or denaturants on recognition expand their use in the case of poorly soluble targets. The ligand′s synthetic nature overcomes all the problems associated with the use of immunoaffinity columns, since the possibility of biological contaminations is extremely limited. Numerous examples demonstrate the usefulness of this methodology, which allows the creation of a large number of peptidyl ligands tailored to specific purification needs. Recognition properties of complementary peptides can be applied also to the development of solid-phase assays for the identification and quantification of different molecular targets. As antibody mimetics, they can be used on solid phases to capture, and then to detect and consequently quantify, the desired target polypeptide in complex biological mixtures. Even if assay sensitivity cannot be compared with conventional antibody-based assays, the simplicity of complementary peptide design and production makes their use an attractive alternative in various circumstances.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 121-129"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Peptide Design Using a Genetically Patterned Binary Code: Growth Hormone-Releasing Hormone as a Model","authors":"Weigent Douglas A.,&nbsp;Clarke Benjamin L.,&nbsp;Blalock J.Edwin","doi":"10.1006/immu.1994.1042","DOIUrl":"10.1006/immu.1994.1042","url":null,"abstract":"<div><p>This paper reviews a method for the design of peptides and proteins of predefined structure and function and provides an example. Specifically, an analog of rat growth hormone-releasing hormone (GHRH) (residues 1-23) was synthesized by solid-phase methods based on a reversed sequence of the mRNA for GHRH (1-23). The new peptide, designated GHRH 3′-5′, had a hydropathic profile similar to that of native GHRH 5′-3′ (GHRH) but had only 17% primary sequence homology. GHRH 3′-5′ specifically bound to the GHRH receptor on rat pituitary cells and to polyclonal anti-GHRH antibody in ELISA and RIA procedures. Additionally, GHRH 3′-5′ blocked the <em>in vitro</em> stimulation of GH RNA synthesis and <em>in vitro</em> and <em>in vivo</em> GH release mediated by GHRH. These data show that 3′-5′ GHRH with little sequence homology to native rat GHRH is an antagonist and further supports the importance of the linear pattern of hydropathy to the gross secondary and/or tertiary structure and rudimentary function of peptides and proteins. The impact of these findings on the interaction of complementary peptides is discussed.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 91-97"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Interactive Determinants on Idiotypic-Anti-idiotypic Antibodies through Comparison of Their Hydropathic Profiles","authors":"Maier Curtis C.,&nbsp;Moseley Hunter N.B.,&nbsp;Zhou Shan-Ren,&nbsp;Whitaker John N.,&nbsp;Blalock J.Edwin","doi":"10.1006/immu.1994.1044","DOIUrl":"10.1006/immu.1994.1044","url":null,"abstract":"<div><p>We have written a computer program to aid in the identification of interaction sites between proteins. The program compares the hydropathic profiles of the two interacting proteins and reports sites, demonstrating an exact pattern of inverted hydropathy. If these regions are surface accessible in the folded proteins, they are considered putative binding or docking sites and can be tested as such. In this report, we apply this program to the localization of residues involved in the anti-idiotope of a monoclonal antibody (mAb), F30C7. The anti-idiotope of F30C7 partially resembles the structure of the peptide antigen, human myelin basic protein (MBP) acetyl 1-9, used to elicit the idiotope bearing mAbs (Ab1). The sequences of F30C7 variable regions are compared to the variable regions of Ab1, as well as to the peptide antigen used to elicit F30C7. Sites of hydropathic cornplementarity In F30C7 with Ab1 that also have sequential homology with MBP 1-9 were located, and a synthetic peptide designed from these sequences was found to structurally resemble MBP 1-9 in that it: (i) inhibited Ab1 binding to MBP 1-9 and (ii) partially inhibited the binding of F30C7 to Ab1. Thus the portion of the anti-idiotope of F30C7 resembling MBP 1-9 was determined with the aid of this program. Other hits between F30C7 and Ab1 also occurred, and future studies will determine whether or not these sites might further contribute to the anti-idiotope.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 107-113"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1044","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibodies to Complementary Peptides as Probes for Receptors","authors":"McGuigan James E.","doi":"10.1006/immu.1994.1050","DOIUrl":"10.1006/immu.1994.1050","url":null,"abstract":"<div><p>Peptide hormones initiate their physiological responses by binding to receptor proteins embedded in the plasma membranes of their target cells. Mechanisms accounting for specific protein-protein interactions, such as peptide hormone binding by cell receptors or epitope binding by antibody have not been defined. A fundamental tenet of the immunological network hypothesis is the generation of anti-idiotypic antibodies to epitopes located in the hypervariable regions of antibody evoked in the same animal species. Anti-idiotypic antibodies to antibodies to peptide hormones with specificity for epitopes involving antibody binding sites may mimic the actions of the peptide hormone by binding to receptors and evoke cell responses associated with the hormone. A provocative relationship was identified in the genetic code, which recognized that complementary codons for strongly hydrophobic amino acids code for strongly hydrophilic amino acids. This led to the proposal and then to demonstration that peptide pairs based on the nucleotide sequences of complementary codons bind one another. It was then proposed that immunization with complementary peptides to peptide hormones may produce antibodies which, analogous to anti-idiotypic antibodies, may mimic the hormone. Some antibodies to complementary peptides for peptide hormones have been shown to mimic the peptide hormones by binding to their receptors and evoking cell responses characteristic of those of the hormones. Exploiting these relationships, some antibodies to complementary peptides for peptide hormones have been used to identify, purify, and characterize receptor proteins for peptide hormones. Polypeptide hormones initiate their characteristic physiologic effects by binding to specific receptor proteins located on the plasma membranes of their target cells. Cell receptor proteins for peptide hormones vary in their structural characteristics, in their mechanisms for signal transduction, and in their affinity and specificity for their peptide ligands. Receptor proteins have been classified based on their transmembrane-spanning segments, identified by defined consecutive groupings of hydrophobic and hydrophilic amino acids and the signal transduction mechanisms by which they evoke the intracellular events leading to physiological responses characteristically identified with the hormone. As for other proteins, the functions of receptor proteins, including the specificity and affinity of their ligand binding, are dictated by their primary amino acid sequence structures. Receptor proteins for peptide hormones in the plasma membrane are relatively few in number and undergo a series of intracellular trafficking steps following hormone binding. Protein-protein interactions are clearly of major biological importance and have been the subject of intensive investigation. These include interactions of antibody binding sites with their epitopes, proteolytic enzymes with their substrates, and receptor prot","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 158-166"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of Complementary Peptides and Their Antibodies in B-Cell-Mediated Autoimmune Disease: Prevention of Experimental Autoimmune Myasthenia Gravis with a Peptide Vaccine","authors":"Araga Shigeru,&nbsp;Blalock J.Edwin","doi":"10.1006/immu.1994.1047","DOIUrl":"10.1006/immu.1994.1047","url":null,"abstract":"<div><p>We have developed and describe a new method of altering B-cell-mediated autoimmune diseases by induction of anti-idiotypic (Id) antibodies (Abs) by immunization with complementary peptides. Specifically, a peptide denoted RhCA 67-16 encoded by RNA complementary to RNA for the <em>Torpedo</em> acetylcholine receptor (AChR) main immunogenic region, AChR 61-76, was tested in the Lewis rat model of experimental autoimmune myasthenia gravis (EAMG). Immunization with RhCA 67-16 induced monoclonal and polyclonal anti-Id Ab against Abs to <em>Torpedo</em> AChR 61-76. RhCA 67-16 antisera inhibited AChR binding by AChR-specific Abs. In addition, a mAb to RhCA 67-16 (denoted TCM 240) recognized two well known EAMG-causing mAbs, 6 and 35. TCM 240, but not a control mAb F28C, inhibited mAb 6 binding to <em>Torpedo</em> AChR 67-76 peptide. mAb 35 binding to TCM 240 was inhibited by native <em>Torpedo</em> AChR as well as by RhCA 67-16. In <em>in vivo</em> experiments, immunization with a RhCA 67-16 keyhole limpet hemacyanin (KLH) conjugate blocked the development of EAMG after challenge with native <em>Torpedo</em> AChR (25% disease incidence versus 90% in the controls). This new approach may provide a novel therapy for MG and perhaps other B-cell-mediated autoimmune disorders through the induction of anti-Id Abs with complementary peptide antigens.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 130-135"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1047","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Complementary Peptides That Interfere with Platelet Aggregation and Adherence","authors":"Gartner T.Kent,&nbsp;Taylor Donald B.,&nbsp;Derrick Jerry","doi":"10.1006/immu.1994.1049","DOIUrl":"10.1006/immu.1994.1049","url":null,"abstract":"<div><p>This article describes the application of the molecular recognition hypothesis to the critically important process of fibrinogen binding to platelets, a process that is the subject of extensive and intensive basic and clinical research. The objectives of the studies summarized below were to design, synthesize, and characterize peptides that can inhibit the binding of fibrinogen and related ligands to human platelets and thereby prevent platelet aggregation, adhesion, and clot retraction. The purpose of doing this work was twofold: first, to determine whether the molecular recognition hypothesis could serve as a useful rationale for the design of peptides that can specifically inhibit the binding of fibrinogen and related ligands to platelets; and second, to use these peptides to try to learn where fibrinogen binds to the platelet fibrinogen receptor. It was hoped that the results obtained not only would provide insight into platelet function but also might provide a rationale for the design of a clinically useful anti-thrombotic agent. Although our studies are not complete, they have resulted in the design of a variety of peptides that can inhibit platelet aggregation, adhesion, and clot retraction as a consequence of specifically inhibiting the binding of fibrinogen and related ligands to the platelet fibrinogen receptor. Although none of these peptides appears to be ligand specific, one or two of them may be specific for platelets .</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 148-157"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Monoclonal Autoantibody against a Complementary Peptide Recognizes Interstitial Collagenase","authors":"de Souza Sandro José,&nbsp;Madaio Michael P.,&nbsp;Neto Luis Juliano,&nbsp;Brentani Ricardo Renzo","doi":"10.1006/immu.1994.1052","DOIUrl":"10.1006/immu.1994.1052","url":null,"abstract":"<div><p>A monoclonal autoantibody (mab 16) against the complementary peptide TKKTLRT, which was deduced from the collagenase-sensitive site in collagen, is described. Mab 16 recognized interstitial collagenase as visualized by ELISA and immunoprecipitation assays. Moreover, mab 16 was able to inhibit partially the collagenolytic activity of keratinocyte supernatant. Finally, it was possible to immunopurify collagenase using a mab 16/Sepharose column.</p></div>","PeriodicalId":79341,"journal":{"name":"ImmunoMethods","volume":"5 2","pages":"Pages 172-176"},"PeriodicalIF":0.0,"publicationDate":"1994-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/immu.1994.1052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18873599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}