Unknown

Dataset Information

0

Mutational analysis of the proteolytic domain of pregnancy-associated plasma protein-A (PAPP-A): classification as a metzincin.


ABSTRACT: The bioavailability of insulin-like growth factor (IGF)-I and -II is controlled by six IGF-binding proteins (IGFBPs 1-6). Bound IGF is not active, but proteolytic cleavage of the binding protein causes release of IGF. Pregnancy-associated plasma protein-A (PAPP-A) has recently been found to cleave IGFBP-4 in an IGF-dependent manner. To experimentally support the hypothesis that PAPP-A belongs to the metzincin superfamily of metalloproteinases, all containing the elongated zinc-binding motif HEXXHXXGXXH (His-482-His-492 in PAPP-A), we expressed mutants of PAPP-A in mammalian cells. Substitution of Glu-483 with Ala causes a complete loss of activity, defining this motif as part of the active site of PAPP-A. Interestingly, a mutant with Glu-483 replaced by Gln shows residual activity. Known metzincin structures contain a so-called Met-turn, whose strictly conserved Met residue is thought to interact directly with residues of the active site. By further mutagenesis we provide experimental evidence that Met-556 of PAPP-A, 63 residues from the zinc-binding motif, is located in a Met-turn of PAPP-A. Our hypothesis is also supported by secondary-structure prediction, and the ability of a 55-residue deletion mutant (d[S498-Y552]) to express and retain antigenecity. However, because PAPP-A differs in the features defining the individual established metzincin families, we suggest that PAPP-A belongs to a separate family. We also found that PAPP-A can undergo autocleavage, and that autocleaved PAPP-A is inactive. A lack of unifying elements in the sequences around the found cleavage sites of PAPP-A and a variant suggests steric regulation of substrate specificity.

SUBMITTER: Boldt HB 

PROVIDER: S-EPMC1222068 | biostudies-other | 2001 Sep

REPOSITORIES: biostudies-other

Similar Datasets

| S-EPMC4571946 | biostudies-literature
| S-EPMC3617297 | biostudies-literature
| S-EPMC6744435 | biostudies-literature
| S-EPMC3375443 | biostudies-literature
| S-EPMC8024009 | biostudies-literature
| S-EPMC8700087 | biostudies-literature
| S-EPMC8805841 | biostudies-literature
| S-EPMC177181 | biostudies-other
| S-EPMC1219474 | biostudies-other
| S-EPMC4198481 | biostudies-literature