"IZ3272"
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Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
Přeruš. str. : il. ; 32 cm
Projekt vyhledávání a analýzy ligandů proteinázy HIV technikou expozice na bakteriofágu navrhuje konstrukci protilátkových fragmentů Fv, mutační analýzu existujících protilátek a konstrukci náhodných peptidů k inhibici sekundárních funkčních míst enzymu.
- MeSH
- HIV genetika MeSH
- klonování DNA MeSH
- krystalografie rentgenová metody MeSH
- mapování epitopu MeSH
- monoklonální protilátky MeSH
- mutační analýza DNA MeSH
- rekombinantní proteiny MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- biologie
- dermatovenerologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu IGA MZ ČR
F11.2.32, a monoclonal antibody raised against HIV-1 protease (Kd = 5 nM), which inhibits proteolytic activity of the enzyme (K(inh) = 35(+/-3)nM), has been studied by crystallographic methods. The three-dimensional structure of the complex between the Fab fragment and a synthetic peptide, spanning residues 36 to 46 of the protease, has been determined at 2.2 A resolution, and that of the Fab in the free state has been determined at 2.6 A resolution. The refined model of the complex reveals ten well-ordered residues of the peptide (P36 to P45) bound in a hydrophobic cavity at the centre of the antigen-binding site. The peptide adopts a beta hairpin-like structure in which residues P38 to P42 form a type II beta-turn conformation. An intermolecular antiparallel beta-sheet is formed between the peptide and the CDR3-H loop of the antibody; additional polar interactions occur between main-chain atoms of the peptide and hydroxyl groups from tyrosine residues protruding from CDR1-L and CDR3-H. Three water molecules, located at the antigen-antibody interface, mediate polar interactions between the peptide and the most buried hypervariable loops, CDR3-L and CDR1-H. A comparison between the free and complexed Fab fragments shows that significant conformational changes occur in the long hypervariable regions, CDR1-L and CDR3-H, upon binding the peptide. The conformation of the bound peptide, which shows no overall structural similarity to the corresponding segment in HIV-1 protease, suggests that F11.2.32 might inhibit proteolysis by distorting the native structure of the enzyme.
- MeSH
- HIV-proteasa MeSH
- hybridomy MeSH
- imunoglobuliny - Fab fragmenty * genetika chemie MeSH
- inhibitory HIV-proteasy * chemie MeSH
- klonování DNA MeSH
- krystalografie rentgenová MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- monoklonální protilátky * genetika chemie MeSH
- počítačová simulace MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- vazebná místa MeSH
- zkřížené reakce MeSH
- Publikační typ
- práce podpořená grantem MeSH
F11.2.32, a monoclonal antibody directed against the HIV-1 protease, displays strong inhibitory effects toward the catalytic activity of the enzyme. The antibody cross-reacts with peptides 36-46 and 36-57 from the protease. Crystals of the Fab have been obtained both in the free state and as complexes formed with the protease peptide fragments, 36-46 and 36-57. Diffraction data have been collected for the free and complexed forms of Fab F11.2.32 and preliminary models for the crystal structures were obtained by molecular replacement.
- MeSH
- HIV antigeny * imunologie MeSH
- HIV protilátky * farmakologie chemie imunologie MeSH
- HIV-1 * enzymologie imunologie MeSH
- HIV-proteasa * imunologie MeSH
- imunoglobuliny - Fab fragmenty chemie imunologie MeSH
- inhibitory HIV-proteasy * chemie imunologie MeSH
- krystalografie rentgenová MeSH
- monoklonální protilátky * farmakologie chemie imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- peptidové fragmenty imunologie MeSH
- rekombinantní fúzní proteiny imunologie MeSH
- zkřížené reakce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
