2001 Volume 130 Issue 2 Pages 235-241
The genomic organization of the gene encoding the human adipocyte-derived leucine aminopeptidase (A-LAP) has been determined. The gene is composed of 20 exons and 19 introns and spans approximately 47 kilobases of chromosome 5q15. The gluzincin aminopeptidase motif, the HEXXH(X)18E zinc-binding motif essential for enzymatic activity, is encoded by exons 6 and 7. A comparison of the exon/intron boundaries, together with phylogenetic analysis, shows the close relationship between A-LAP and placental leucine aminopeptidase (P-LAP)/oxylocinase, another gluzincin aminopeptidase considered to be important for the maintenance of normal pregnancy. Primer extension analysis revealed two transcriptional initiation sites. Analysis of the sequence immediately upstream of the transcriptional initiation sites revealed that the A-LAP promoter contains no canonical TATA-or CCAAT-box, but has a PyPyA+1N(T/A)PyPy initiator consensus sequence and multiple putative regulatory elements. Finally, luciferase-reporter assays revealed a functional promoter activitiy of the 5'-flanking region of the gene, and suggested that the activity is regulated in a cell type-specific manner.