title: Evolutionary and biochemical differences between human and monkey acidic mammalian chitinases.
abstract: Acidic mammalian chitinase (AMCase), an enzyme implicated in the pathology of asthma, is capable of chitin cleavage at a low pH optimum. The corresponding gene (CHIA) can be found in genome databases of a variety of mammals, but the enzyme properties of only the human and mouse proteins were extensively studied. We wanted to compare enzymes of closely related species, such as humans and macaques. In our attempt to study macaque AMCase, we searched for CHIA-like genes in human and macaque genomes. We found that both genomes contain several additional CHIA-like sequences. In humans, CHIA-L1 (hCHIA-L1) is an apparent pseudogene and has the highest homology to CHIA. To determine which of the two genes is functional in monkeys, we assessed their tissue expression levels. In our experiments, CHIA-L1 expression was not detected in human stomach tissue, while CHIA was expressed at high levels. However, in the cynomolgus macaque stomach tissue, the expression pattern of these two genes was reversed: CHIA-L1 was expressed at high levels and CHIA was undetectable. We hypothesized that in macaques CHIA-L1 (mCHIA-L1), and not CHIA, is a gene encoding an acidic chitinase, and cloned it, using the sequence of human CHIA-L1 as a guide for the primer design. We named the new enzyme MACase (Macaca Acidic Chitinase) to emphasize its differences from AMCase. MACase shares a similar tissue expression pattern and pH optimum with human AMCase, but is 50 times more active in our enzymatic activity assay. DNA sequence of the mCHIA-L1 has higher percentage identity to the human pseudogene hCHIA-L1 (91.7%) than to hCHIA (84%). Our results suggest alternate evolutionary paths for human and monkey acidic chitinases.
authors.affiliation: Pfizer Biotherapeutics Research and Development, Department of Inflammation and Immunology, 200 CambridgePark Drive, Cambridge, MA 02140, USA. rkrykbaev@wyeth.com
authors.forename: Rustem
authors.initials: R
authors.lastname: Krykbaev
authors.forename: Lori J
authors.initials: LJ
authors.lastname: Fitz
authors.forename: Padmalatha S
authors.initials: PS
authors.lastname: Reddy
authors.forename: Aaron
authors.initials: A
authors.lastname: Winkler
authors.forename: Dejun
authors.initials: D
authors.lastname: Xuan
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Yang
authors.forename: Margaret
authors.initials: M
authors.lastname: Fleming
authors.forename: Stanley F
authors.initials: SF
authors.lastname: Wolf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017124:Conserved Sequence; D030541:Databases, Genetic; D019143:Evolution, Molecular; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008252:Macaca fascicularis / Q000235:genetics*; D008253:Macaca mulatta / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016415:Sequence Alignment
chemical_list: D002686:Chitin; D002688:Chitinases