Cloning and characterization of a novel peptidase from rat and human ileum.
A novel 100-kDa ileal brush border membrane protein (I100) has been purified by anionic glycocholate affinity chromatography. Polyclonal antibodies raised against this protein were utilized to clone and characterize I100 in rats. A partial length human I100 cDNA was identified by hybridization screening. In the rat, the I100 protein is a 746-amino acid glycosylated (calculated core molecular mass of 80 kDa) type II integral membrane protein found on the apical surface of ileal villus enterocytes. Its 2.6-kilobase mRNA is expressed in distal small intestine in rats and in humans. The I100 cDNA is homologous to but distinct from human prostate-specific membrane antigen and rat brain N-acetylaspartylglutamate peptidase. It is expressed on both the basolateral and apical surfaces of stably transfected Madin Darby canine kidney cells. Analysis of these stably transfected Madin Darby canine kidney cells and I100 immunoprecipitates of rat ileal brush border membrane vesicles reveals that it has dipeptidyl peptidase IV activity. Future invesitgations will need to determine the exact substrate specificity of this novel peptidase.
Shneider
B L
BL
Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Thevananther
S
S
Moyer
M S
MS
Walters
H C
HC
Rinaldo
P
P
Devarajan
P
P
Sun
A Q
AQ
Dawson
P A
PA
Ananthanarayanan
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D004285:Dogs; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007082:Ileum / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016415:Sequence Alignment; D014018:Tissue Distribution; D014162:Transfection
D012333:RNA, Messenger; D018819:Dipeptidyl Peptidase 4