Physiochemical and immunological comparisons between angiotensin I-converting enzymes purified from different mammalian species.
Angiotensin I-converting enzyme (ACE) was purified from lungs of pig, rat, monkey and human for comparison of its physicochemical, enzymatic and immunological properties. The protocol involved three chromatographic steps after detergent extraction, i.e. DEAE-Sphérodex ion exchange, lisinopril-Sepharose affinity and Superose 12 HPLC, plus Mono-Q HPLC for monkey ACE. Purified ACE's presented numerous homologies: in particular, closely similar specific activities, catalytic efficiencies, Km's, optimal pH and chloride activations; the molecular weights were about 170 kDa by SDS-PAGE and 320 kDa by gel-filtration on Superose 12; the isoelectric points were about 4.5-4.7. Specific polyclonal antibodies recognized the antigen (porcine ACE) as well as rat, monkey and human ACEs. In contrast, three monoclonal antibodies (F02.4.1, F01.1.3 and F03) produced against porcine ACE showed some differences: they only reacted with pig enzyme and only one (F0.2.4.1) was anticatalytic. Moreover, the cross-reactivity judged on ELISA with porcine ACE characterized different epitopes specific for the porcine enzyme. In particular, the binding of F02.4.1 was not diminished by previous treatment with saturating concentrations of synthetic competitive ACE inhibitors. Thus, the extrapolation to human of data obtained on animal models should be possible at least for pharmacological and medical trials.
Bénéteau-Burnat
B
B
Laboratoire de Biochimie A, Hôpital Saint-Antoine, Paris, France.
Tahraoui
A
A
Barbut
F
F
Giboudeau
J
J
Baudin
B
B
D000818:Animals; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000882:Haplorhini; D006801:Humans; D008168:Lung / Q000201:enzymology*; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D051381:Rats; D013552:Swine
D000911:Antibodies, Monoclonal; D007703:Peptidyl-Dipeptidase A