PMC

Predicted hydrophobic contacts surrounding V246 in FF8. Based on the enzyme structure, residue A246 (A) and the modeled V246 (B) are colored blue and cyan, respectively, while the surrounding hydrophobic residues are shown in gray. Replacement of A246 with valine might result in better hydrophobic packing. This figure was displayed by using GRASP (20).

Experiments with wild-type (GAAA) or modified (GAAAAA) tetraloops to show that pyrene fluorescence reports on P4–P6 tertiary folding for the T8 tether at A246, U247, U249 and U253. See text for details.

Equilibrium fluorescence titrations of P4–P6 derivatives labeled with pyrene at U247, U249, U253, A246, A256 or C240. For each of the five tethers, the fluorescence spectra were qualitatively similar to those for U107 that are depicted in Figure 4 (data not shown). See Supplementary Data for [Mg²⁺]_1/2 values.

Type I collagenase activity is inhibited in human aortic fibrous caps by the fibroblast activation protein-blocking antibody A246. (A) Movat staining of a fibrous cap in human aortic plaque. The region of interest (black box) is shown at higher magnification in an adjacent section stained for fibroblast activation protein (red), type I collagen (green), and overlay (DAPI = blue; bar = 150 µm). (B) Confocal images of in situ zymography show fibroblast activation protein (red) and cleaved DQ type I collagen (green) in fibrous caps shown by Movat (A) of aortic plaque treated with a control IgG antibody or neutralizing antibody A246 (bar = 10 μm). (C) The graph reveals a significant reduction of cleaved type I collagen colocalized with fibroblast activation protein expression by in situ zymography (n = 10/group). (D) Type I collagenase activity of recombinant human fibroblast activation protein and the neutralizing capacity of A246 was demonstrated by incubation of native human type I collagen with recombinant human fibroblast activation protein in the presence of inhibiting or isotype control antibodies (lanes: 1, rhuFAP; 2, collagen; 3, rhuFAP + collagen; 4, rhuFAP + collagen + A246; 5, rhuFAP + collagen + isotype control antibody; 6, molecular weight marker).