N. Bruneau et al., The affinity binding sites of pancreatic bile salt-dependent lipase in pancreatic and intestinal tissues, J HIST CYTO, 48(2), 2000, pp. 267-276
In previous studies, we have shown that the bile salt-dependent lipase (BSD
L) associates with the Grp94 molecular chaperone, an association that appea
rs to play essential roles in the folding of BSDL. More recently, combined
biochemical and immunocytochemical investigations were carried out to show
that the transport of BSDL occurs via an association with the Grp94 all alo
ng the pancreatic secretory route (ER-Golgi-granules). The Grp94-BSDL compl
ex is secreted with the pancreatic juice into the acinar lumen and reaches
the duodenal lumen, where it is internalized by enterocytes. The dissociati
on of the complex could take place within the endosomal compartment because
BSDL continues further on its way to the basolateral membrane of the enter
ocyte. To localize the affinity binding sites of pancreatic BSDL in pancrea
tic and duodenal tissues, we have used an affinity-gold ultrastructural tec
hnique. BSDL coupled to gold particles appears to interact with specific si
tes in tissue sections. This was confirmed by another indirect morphologica
l approach using biotin-labeled BSDL and streptavidin-gold complexes on tis
sue sections. We have shown that BSDL associates with sites in the pancreat
ic secretory pathway compartments and in the microvilli, the endosomal comp
artment, and the basolateral membrane of enterocytes. By biochemical approa
ches, biotin-labeled BSDL displayed affinities with proteins of 180-190 kD
in both pancreatic and duodenal tissues. We have also shown that the Grp94-
BSDL complexes, which are insensitive to denaturing conditions, are present
in pancreatic homogenate but not in duodenal lysate. Thus, BSDL is able to
bind protein complexes formed by either BSDL-Grp94 or Grp94 dimers.