PURIFICATION AND AMINO-ACID-SEQUENCE OF MOTILIN FROM CAT SMALL-INTESTINE

被引:20
作者
DEPOORTERE, I [1 ]
PEETERS, TL [1 ]
VANDERMEERS, A [1 ]
VANDERMEERSPIRET, MC [1 ]
CHRISTOPHE, J [1 ]
VANTRAPPEN, G [1 ]
机构
[1] ULB,CHIM BIOL LAB,B-1070 BRUSSELS,BELGIUM
关键词
MOTILIN; PURIFICATION; AMINO ACID SEQUENCE; SMALL INTESTINE; CAT;
D O I
10.1016/0167-0115(93)90380-Q
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Motilin was isolated from cat small intestine by a series of chromatographic steps. Using a radioreceptor assay, based upon binding of iodinated porcine motilin to rabbit antral smooth muscle membranes, it was shown that cat duodenal mucosa contains about 495 ng/g tissue, the jejunal mucosa 161 ng/g tissue and the ileal mucosa 95 ng/g tissue motilin. The duodenal mucosa was extracted with 6% acetic acid and concentrated on a cation exchange Whatman CM-52 gel. After lyophilization the material was further purified by gel filtration (Sephadex G-50), followed by reverse phase (C18), cation exchange HPLC (Mono S) and three runs on a reverse phase HPLC (Nucleosil 300-5C18) column. The UV absorbance and the radioreceptor assay were used to monitor the purification. After Mono S chromatography two forms of motilin were detected. The major peak corresponded to a 22 amino acid peptide, which differed only from canine motilin at position 12, where Lys is replaced by Arg. The smaller peak probably corresponds to a deamidated form of this peptide. The sequence homology between cat and porcine/human motilin or cat and rabbit motilin is 81.8% and 72.7%, respectively. The conservation of the first six amino acids in all five species studied is striking, confirming that the biological acitivity of the peptide resides in the N-terminal part.
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收藏
页码:25 / 32
页数:8
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