DOMINANT B-CELL EPITOPE FROM THE PRES2 REGION OF HEPATITIS B VIRUS IN THE FORM OF AN INSERTED PEPTIDE SEGMENT IN MALTODEXTRIN-BINDING PROTEIN

Crystal structure of a dominant B-cell epitope from the preS2 region of hepatitis B virus in the form of an inserted peptide segment in maltodextrin-binding protein.

Maltodextrin-Binding Protein Hybrids Carrying Epitopes from the Pres2 Region of Hepatitis B Virus: Expression, Antibody-Binding and Preliminary Crystallographic Studies

PROTEIN CONCENTRATION 3.0MG/ML, BUFFER 0.1M TRIS-HCL PH 8.5 WITH 0.2 M MGCL2, 1 MM MALTOSE, 26% PEG 4000.

MALE-B363 IS AN INSERTION/DELETION MUTANT OF THE E. COLI MALTODEXTRIN-BINDING PROTEIN (MBP) CARRYING THE AMINO-ACID SEQUENCE OF A B-CELL EPITOPE FROM THE PRES2 REGION OF THE HEPATITIS B VIRUS SURFACE ANTIGEN. AMINO ACIDS BELONGING TO THE PRES2 EPITOPE ARE DESIGNATED 132E TO 145E (POSITIONS 132-145 OF THE VIRAL SURFACE ANTIGEN). RESIDUES DESIGNATED 364I, 365I, AND 380I-389I, WHICH FLANK THE EPITOPE SEGMENT, RESULT FROM THE GENETIC CONSTRUCTION USED TO PERMIT THE INSERTION. NUMBERING OF THE INSERTED SEQUENCE IS THUS 364I, 365I, 132E TO 145E, AND 380I TO 389I. THESE RESIDUES REPLACE WILD-TYPE MBP RESIDUES FROM POSITIONS 364 - 370. THE NUMBERING OF MBP RESIDUES IN THIS ENTRY CORRESPONDS TO THE SEQUENCE POSITIONS OF WILD-TYPE MBP.

THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT CONTAINS THREE INDEPENDENT MOLECULES OF MALE-B363. EACH IS ASSOCIATED WITH A BOUND MOLECULE OF MALTOSE SUBSTRATE. THE FOLLOWING CHAIN IDENTIFIERS ARE USED: CHAIN A, MALE-B363 MOLECULE 1; CHAIN B, MALE-B363 MOLECULE 2; CHAIN C, MALE-B363 MOLECULE 3; CHAIN D, HET GROUP ASSOCIATED WITH MOLECULE 1; CHAIN E, HET GROUP ASSOCIATED WITH MOLECULE 2; CHAIN F, HET GROUP ASSOCIATED WITH MOLECULE 3; CHAIN X, SOLVENTS ASSOCIATED WITH MOLECULE 1; CHAIN Y, SOLVENTS ASSOCIATED WITH MOLECULE 2; CHAIN Z, SOLVENTS ASSOCIATED WITH MOLECULE 3.

TRANSPORT, PRES2, EPITOPE, HEPATITIS B, MALTOSE-BINDING PROTEIN