Functional comparison of recombinant acidic mammalian chitinase with enzyme from murine bronchoalveolar lavage.
Acidic mammalian chitinase (AMCase) is an enzyme that selectively degrades the biopolymer chitin. Several chitinase enzymes are utilized by mammals to hydrolyze chitin encountered by inhalation and ingestion. AMCase is distinct from other mammalian chitinases as its activity is retained in strongly acidic conditions (pH <2.0). AMCase expression is induced by antigen-induced mouse models of allergic lung inflammation. This protein has also been implicated in the pathogenesis of asthma although its precise role is poorly defined. We describe a novel way to express and purify active murine AMCase. This material retains properties observed in mouse bronchoalveolar lavage (BAL) fluid with regard to pH preference of activity and its inhibition by cyclic peptide inhibitors argifin and argadin. We found that chitinase in BAL from both antigen-challenged and control animals have similar properties in this regard. This strongly supports the notion the same enzyme (AMCase) gives rise to chitinase activity in both challenged and unchallenged animals. We also describe expression of active human AMCase. The methods described in this paper provide a reliable source of recombinant AMCase that can be utilized to expand understanding of AMCase's role in regulating allergic inflammation.
Goedken
Eric R
ER
Abbott Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA. eric.goedken@abbott.com
O'Brien
Robert F
RF
Xiang
Tao
T
Banach
David L
DL
Marchie
Susan C
SC
Barlow
Eve H
EH
Hubbard
Stacy
S
Mankovich
John A
JA
Jiang
Jianjun
J
Richardson
Paul L
PL
Cuff
Carolyn A
CA
Cherniack
Andrew D
AD
D000595:Amino Acid Sequence; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid; D019556:COS Cells; D002460:Cell Line; D002688:Chitinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D010456:Peptides, Cyclic; D011994:Recombinant Proteins; C419444:argadin; C413566:argifin; C499255:AMCase, mouse; C499254:CHIA protein, human; D002688:Chitinases