Figure 2.

Identification of a missense mutation in BMP1 in individuals with AR-OI. A: Exon–intron structure of the two alternative spliced isoforms (BMP1 and mTLD) transcribed from the BMP1 locus. B: Sequence analysis of genomic DNA from a control (ctrl), a heterozygous individual (ht), and one of the patients (p) showing the c.747C>G transversion in BMP1 exon 6 (red and squared nucleotide). C: Multiple sequence alignment of a fragment from the catalytic domain of different astacin-like metalloproteases from diverse organisms showing conservation of the F249 residue labeled in red. The lower cluster of sequences corresponds to protein members of the BMP1/tolloid-like subgroup of astacin-like metalloproteases both from vertebrate and invertebrate organisms. Aaa_Asta, astacin from the crayfish A. Astacus (P07584), Tpa_My01 myosinase from giant squid Todarodes pacificus (Q8IU46), Cel_Nas13 metalloproteinase from Caenorhabditis elegans, (Q20191), Qu_Cam-I from quail (AAA20842.1), Hu_Ovast, human ovastacin (Q6HA08), Hu_MEPα (Q16819) and Hu_MEPβ (Q16820), human meprins α and β, Hvu_Hmp1 (AAA92361.2) and Hvu_Hmp2 (AAD33860.1) from Hydra vulgaris, Spu_Span from sea urchin (P98068), Lpo_Astm from the horseshoe crab Limulus polyphemus (B4F320), Ola_Hce1 (NP_001188427.1) and Ola_Lce1 (NP_001098292.1) from medaka fish Oryzias latipes, Dr_tld, Drosophila tolloid (NP_524487.2), Ate_tll, tolloid-like peptidase from the spider Achaearanea tepidariorum (Q75UQ6), Ci_Bmp1 from Ciona intestinalis (NP_001071840.1), Da_Bmp1a from Danio rerio (NP_001035126.1), Xe_Bmp1 from Xenopus laevis (AAI70427.1), Mu_Bmp1 from Mus musculus (NP_033885.2), Hu_BMP1/mTLD, Hu_TLL1 and Hu_TLL2 human BMP1/TLD-like proteases (NP_001190.1/NP_006120.1, NP_036596.3 and NP_036597.1). For most of the proteins shown here, a wider sequence alignment of the catalytic domain and information about substrate specificity have been described previously [Bond and Beynon, 1995; Guevara et al., 2010].