CHROMOGRANIN-B - ISOLATION FROM PHEOCHROMOCYTOMA, N-TERMINAL SEQUENCE, TISSUE DISTRIBUTION AND SECRETORY VESICLE PROCESSING

被引:23
作者
GILL, BM
BARBOSA, JA
DINH, TQ
GARROD, S
OCONNOR, DT
机构
[1] UNIV CALIF SAN DIEGO,DEPT MED,SAN DIEGO,CA 92103
[2] UNIV CALIF SAN DIEGO,CTR MOLEC GENET,SAN DIEGO,CA 92103
关键词
SECRETOGRANIN; ADRENAL MEDULLA;
D O I
10.1016/0167-0115(91)90216-4
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The chromogranins/secretogranins are a family of neuroendocrine vesicle secretory proteins. Immunohistology and immunoblotting have suggested that a major soluble protein in human chromaffin granules may be chromogranin B (CgB). We purified from pheochromocytoma chromaffin granules an SDS-PAGE 110-120 kDa protein whose N-terminal sequence matched that previously deduced from a human CgB cDNA. An antibody directed against a synthetic human CgB N-terminal region specifically recognized the CgB N-terminus, though not the chromogranin A (CgA) N-terminus or the CgB C-terminus on immunoblots. An antiserum directed against CgB's C-terminus also visualize CgB but not CgA. By immunoblotting, CgB was a quantitatively major protein in human pheochromocytoma chromaffin granules, but a relatively minor protein in normal bovine adrenal medullary chromaffin granules. In a variety of normal bovine neuroendocrine tissues, the relative abundance of CgB immunoreactivity on immunoblots was: adrenal medulla > anterior pituitary > pancreas > small intestine, hypothalamus. Immunoblotting of neuroendocrine tissues (or their hormone storage vesicle cores) with both anti N-terminal and anti C-terminal CgB antisera suggested bidirectional cleavage or processing of CgB; in the anterior pituitary, a unique 40 kDa C-terminal fragment was observed. Bidirectional CgB cleavage was also suggested on immunoblots of chromaffin tissue from three species (human, bovine, rat). C-terminal processing of CgB was also confirmed by amino acid sequencing of SDS-PAGE-separated, polyvinylidene difluoride membrane-immobilized CgB fragments from phoechromocytoma chromaffin granules. Whether such fragments possess biological activity remains to be investigated.
引用
收藏
页码:223 / 235
页数:13
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