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J. Biol. Chem., Vol. 277, Issue 44, 42028-42033, November 1, 2002

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Characterization of Four Murine Homologs of the Human ov-serpin Monocyte Neutrophil Elastase Inhibitor MNEI (SERPINB1)^*

Charaf Benarafa^§¶, Jessica Cooley, Weilan Zeng, Phillip I. Bird, and Eileen Remold-O'Donnell^§

From the  Center for Blood Research and ^§ Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115 and the  Department of Biochemistry and Molecular Biology, Monash University, Victoria 3800, Australia

The human ov-serpin monocyte neutrophil elastase inhibitor (MNEI) is encoded by a single gene SERPINB1. It is a highly efficient inhibitor of neutrophil granule proteases. Four murine genes with high sequence identity with MNEI were identified and fully sequenced, and these were named EIA, EIB, EIC, and EID. EIA, EIB and EIC showed the same seven-exon gene structure as SERPINB1. However, EIC included an additional, alternatively spliced, exon due to the insertion of an endogenous retrovirus-like sequence. EID lacked several exons and is a pseudogene. Reverse transcriptase-PCR showed that EIA, like MNEI, is expressed at high levels in many tissues. EIB is mainly expressed in brain, and EIC was only expressed as splicing variants unlikely to encode a functional serpin. Upon incubation with serine proteases, EIA formed inhibitory covalent complexes with pancreatic and neutrophil elastases, cathepsin G, proteinase-3, and chymotrypsin, as previously shown for MNEI, whereas EIB was only able to do so with cathepsin G. According to the new serpin nomenclature, the genes encoding EIA, EIB, EIC, and EID will be called Serpinb1, Serpinb1b, Serpinb1c, and Serpinb1-ps1. These data demonstrate that the four murine homologs of MNEI have met different evolutionary fates, and that EIA is the mouse ortholog of MNEI.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF521697, AF521698, AF521699, and AF521700.

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