PHYSICOCHEMICAL AND IMMUNOLOGICAL COMPARISONS BETWEEN ANGIOTENSIN I-CONVERTING ENZYMES PURIFIED FROM DIFFERENT MAMMALIAN-SPECIES

被引:8
作者
BENETEAUBURNAT, B [1 ]
TAHRAOUI, A [1 ]
BARBUT, F [1 ]
GIBOUDEAU, J [1 ]
BAUDIN, B [1 ]
机构
[1] UNIV PARIS 05,BIOCHIM LAB A,F-75006 PARIS,FRANCE
来源
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY | 1994年 / 109卷 / 04期
关键词
ANGIOTENSIN I-CONVERTING ENZYME; PURIFICATION; COMPARATIVE BIOCHEMISTRY; MONOCLONAL ANTIBODIES; EPITOPES; ANIMAL MODELS; ISOELECTRIC FOCUSING; WESTERN-BLOTTING;
D O I
10.1016/0305-0491(94)90125-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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-Spherodex ion exchange, lisinopril-Sepharose affnity 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, K-m'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 Pat, 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 (F02.4.1) 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.
引用
收藏
页码:623 / 635
页数:13
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