Comparative Sequences of Canine and Feline Endothelin-1

Department of Veterinary Pathobiology, University of Illinois at Urbana-Champaign, Urbana, IL 61802, USA.
Veterinary Clinical Pathology (Impact Factor: 1.27). 02/2003; 32(4):188-94. DOI: 10.1111/j.1939-165X.2003.tb00334.x
Source: PubMed


Endothelin-1 (ET-1, "mature ET-1") is a potent vasoconstrictor peptide that is made along with "big ET-1" from its precursor, preproET-1. Increased plasma concentrations of ET-1 and big ET-1 occur with various forms of cardiovascular disease in humans. Our laboratory is investigating plasma endothelins as diagnostic tests of cardiovascular disease in dogs and cats; however, commercial immunoassays designed specifically for use in dogs and cats are limited.
Amino acid sequences of feline and canine big ET-1 were obtained and used to predict antibody cross-reactivity with immunoassay test kits from other species.
Genomic DNA was extracted from peripheral blood and total RNA was extracted from canine and feline left ventricles for reverse transcription polymerase chain reaction (PCR) and PCR amplification of segments of the canine and feline preprohormone containing the big ET-1 sequences. The derived amino acid sequences were compared with known big ET-1 and ET-1 sequences of several other species, including human, mouse, and rat.
Feline and canine big ET-1 had 87-97% and 89-100% homology, respectively, with that of other mammalian species. Canine ET-1 was identical to human, mouse, and rat ET-1. In contrast, the amino acid sequence of feline ET-1 was unique owing to a leucine for methionine substitution at position 7.
It is highly likely that anti-human and anti-rodent ET-1 antibodies will cross-react with mature canine ET-1. In contrast, antibodies to mature ET-1 intended for use with feline tissues and antibodies to big ET-1 in either dogs or cats may have partial or no cross-reactivity depending on the peptide sequences used to produce the antibodies.

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    • "Big ET-1 was measured in all samples using a sandwich ELISA (IBL Hamburg) validated in dogs (Schellenberg et al., 2008). The kit includes a primary capturing antibody against the C-terminal 22–38 amino acid sequence of rat big ET-1, which shows close homology to the canine amino acid sequence (Biondo et al., 2003). The secondary antibody for detection is directed against a different antigenic site of the molecule. "

    Full-text · Article · Sep 2013 · The Veterinary Journal
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    • "similar but not homologous ( Biondo et al . , 2003 ) . Poor cross - reactivity of the anti - Big - ET anti - body with canine Big - ET may be another reason for the poor results obtained with this kit . In contrast , canine and rat amino acid sequences for Big - ET show closer homology at the antibody - binding site than with the human sequences ( Biondo et al . , 2003 ) . Better performance of the rat Big - ET kit could be expected and was confirmed in our study ."
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