Antigenicity and conformational analysis of the Zn2+-binding sites of two Zn2+-metalloproteases: Leishmania gp63 and mammalian endopeptidase-24.11
Φόρτωση...
Ημερομηνία
Συγγραφείς
Soteriadou, K. P.
Tzinia, A. K.
Panou-Pomonis, E.
Tsikaris, V.
Sakarellos-Daitsiotis, M.
Sakarellos, C.
Papapoulou, Y.
Matsas, R.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Portland Press
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
Biochemical Journal
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
The antigenic properties of the Zn2+-binding region of two Zn2+- metalloproteases, Leishmania surface protease gp63 and mammalian endopeptidase-24.11 (E-24.11), possessing in their active site the characteristic amino acid sequence HEXXH, were investigated by using oligoclonal antibodies raised against two synthetic peptides, V(1)VTHEMAHALGL(11) (pepgp63) and V(1)IGHEITHGFD(11) (pepE-24.11), containing the respective Zn2+-binding sites of the cognate protein. The affinity-purified antibodies, tested on synthetic peptides modelled from the active sites of ten different Zn2+-metalloproteases, showed high selectivity for their respective peptides, However, cross-reactivity was revealed when the antibodies were tested against the gp63 and E-24.11 molecules. A panel of synthetic peptide analogues and peptides of various size was synthesized and used for the fine antigenic characterization of pepgp63 and pepE-24.11. The shortest peptides capable of significant antibody binding were the pentapeptides V(1)VTHE(5) and E(5)ITHG(9) for pepgp63 and pepE-24.11 respectively. His(4) and Glu(5) were found to be indispensable for anti-pepgp63 binding to pepgp63, whereas in the case of pepE-24.11, Glu(5) and His(8) were found to be critical. The conformational characteristics of the two peptides correlate well with the observed differences in their antigenicity. H-1-NMR studies showed that pepgp63 adopts a folded structure whereas pepE-24.11 takes up a rather flexible conformation. Moreover, the antigenically critical His(4) of pepgp63 contributes to the structural stabilization of the peptide. Similarly, the antigenically critical Hiss Of pepE-24.11 is involved in partial structural stabilization of its C-terminal region. The generated antibodies may be useful tools for identifying and classifying proteins possessing similar Zn2+-binding motifs and/or environments.
Περιγραφή
Λέξεις-κλειδιά
nuclear-magnetic-resonance, major surface glycoprotein, 24.11 enkephalinase, molecular-dynamics, peptide-synthesis, monoclonal-antibodies, schwann-cells, binding-sites, protein, promastigotes
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
<Go to ISI>://A1996TR39300014
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείας