Molecular cloning and sequencing of the cDNA for rat mesenteric arterial bed elastase-2, an angiotensin II-forming enzyme.
A 28.5-kD protein expressed in rat mesenteric arterial bed (MAB) perfusate with angiotensin II-forming ability was previously characterized. This protein, a member of the elastase-2 family of enzymes, seems to be the only representative of this family of proteases to be secreted outside the digestive tract and implicated in the generation of angiotensin II. The cloning and sequencing of the cDNA for the rat MAB elastase-2 by using reverse transcription polymerase chain reaction are reported. The sequence of this cDNA was found to be identical to the sequence of the rat pancreatic elastase-2; the cDNA is 909 nucleotides in length plus a poly (A) tail and encodes a preproenzyme of 271 amino acids. Analysis of the putative amino acids in the extended angiotensin I binding site of the rat MAB elastase-2 reveals features that could explain the dipeptidyl carboxypeptidase-like activity required for efficient conversion of angiotensin I to angiotensin II. Additionally, the sequence reveals structural features that could contribute to the lack of activity of this enzyme toward angiotensin II. Rat MAB elastase-2 was expressed in mesenteric arteries and lung but not in aorta. These results may also indicate that rat MAB elastase-2 is expressed in resistance vessels but not in conduit vessels.
Santos
Carlos F
CF
University of São Paulo, School of Medicine of Ribeirão Preto, Department of Pharmacology, Brazil.
Oliveira
Eduardo B
EB
Salgado
Maria Cristina O
MC
Greene
Andrew S
AS
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification*; D008297:Male; D008638:Mesenteric Arteries / Q000201:enzymology*; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000706:statistics & numerical data; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; D000804:Angiotensin II; D012697:Serine Endopeptidases; C117309:pancreatic elastase II