Figure 1 : Gp41-MinTT expression and purification.

From: Proteoliposomal formulations of an HIV-1 gp41-based miniprotein elicit a lipid-dependent immunodominant response overlapping the 2F5 binding motif

Figure 1

(a) Schematic representation of gp41, gp41-Min and gp41-MinTT proteins is shown. FP, fusion peptide (blue); HR1, N-terminal heptad repeat (red); S-S, disulfide loop (brown); HR2, C-terminal heptad repeat (green); MPER, membrane proximal external region (yellow); TM, transmembrane domain (purple); CT, cytoplasmic tail (gray); TT, tetanus toxoid epitope (light green); 6xHis, 6-histidine tag (orange). MPER-spanning sequence (residues 659–683, HXB2 numbering) is depicted. MPER sequences containing the 2F5, 4E10 and 10E8 epitopes are underlined. 2F5 neutralizing core and residues equally recognized by 4E10 and 10E8 are highlighted in blue and red respectively. (b) Gel filtration chromatography. Elution profile of the latter purification step is shown. (c) A highly pure 15KDa protein was recovered and concentrated from central fractions of the largest peak shown in panel B (44–49 mL fractions), as confirmed by SDS-PAGE and coomassie staining (left) and by Western blot using the 2F5 antibody (right). Molecular markers are indicated. (d) Antigenicity of purified gp41-MinTT protein determined by ELISA using serial dilutions of D50 (anti-HR2) and 2F5 (anti-MPER) antibodies.