The role of the acid-labile subunit in regulating insulin-like growth factor transport across human umbilical vein endothelial cell monolayers.

ALS significantly regulates IGF transport across endothelial cells, but other factors also play a role.

Summary and findings

This study examined the role of the acid-labile subunit (ALS) in regulating insulin-like growth factor (IGF) transport across human umbilical vein endothelial cell (HUVEC) monolayers. The transport of (125)I-des(1-3)IGF-I was found to be more rapid than (125)I-IGF-I. The findings indicate that ALS plays a significant role in regulating IGF transport, but other factors are also involved.

Abstract

We have established a human umbilical vein endothelial cell (HUVEC) monolayer system to study the role of complex formation with IGF binding protein (IGFBP) and acid-labile subunit (ALS) on the transendothelial transport of IGF. Incubation with recombinant human IGFBP-3 alone did not retard (125)I-IGF-I or -II transport, but addition of ALS caused marked inhibition. Transport of (125)I-des(1-3)IGF-I was more rapid than (125)I-IGF-I, suggesting the presence of some endogenous IGFBPs, although these were undetectable by affinity labeling of cells or medium. In the presence of ALS, recombinant human IGFBP-5 also retarded IGF transport, although significantly less than IGFBP-3, despite their similar ternary complex formation. In contrast, IGFBP-3 mutated in its ALS-binding domain was not inhibitory. To study IGF transport by pregnancy-proteolyzed IGFBP-3, we prepared [Tyr(31)]monoiodoIGF-I, the only iodoIGF-I form that reacts normally with proteolyzed IGFBP-3. In the presence of ALS, IGFBP-3 isolated by immunoaffinity chromatography from second-trimester pregnancy serum significantly retarded IGF transport, but to a lower extent than IGFBP-3 isolated from normal serum, despite normal ALS binding. This study demonstrates the key role of ALS in regulating transendothelial IGF transport, but indicates that other factors are also involved. Our data suggest that pregnancy-proteolyzed IGFBP-3, despite forming normal ternary complexes, is less effective than intact IGFBP-3 in retarding IGF egress from the circulation.

Limitations

Not reported in abstract.

Elsewhere in the IGF-1 DES (Des(1-3) IGF-1) corpus