PROCESSING OF MUTATED PROINSULIN WITH TETRABASIC CLEAVAGE SITES TO BIOACTIVE INSULIN IN THE NONENDOCRINE CELL-LINE, COS-7

被引:44
作者
YANAGITA, M
NAKAYAMA, K
TAKEUCHI, T
机构
[1] GUNMA UNIV,INST ENDOCRINOL,DIV MOLEC ENDOCRINOL,SHOWA MACHI,MAEBASHI,GUNMA 371,JAPAN
[2] UNIV TSUKUBA,CTR GENE EXPT,TSUKUBA,IBARAKI 305,JAPAN
关键词
PROHORMONE PROCESSING; TETRABASIC PROCESSING SITE; PROINSULIN; INSULIN; FURIN; COS-7; CELL;
D O I
10.1016/0014-5793(92)81366-T
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
The amino acid sequence, Arg-4-X-3-Lys/Arg-2-Arg-1 down X+1, is thought to be a concensus processing site for a constitutive secretory pathway in non-endocrine cells. We created a mutant proinsulin DNA with a peptide structure of B chain-Arg-Arg-Lys-Arg-C peptide-Arg-Arg-Lys-Arg-A chain, which compares to the native proinsulin structure of B chain-Arg-Arg-C peptide-Lys-Arg-A chain. When the mutant insulin was expressed in a monkey kidney-derived cell line, COS-7, approximately 60% of the total immunoreactive insulin appeared as mature insulin in the culture medium. This conversion to the mature form was strikingly facilitated by co-expressing the mutant proinsulin with furin, a homologue of the yeast endoprotease, Kex2.
引用
收藏
页码:55 / 59
页数:5
相关论文
共 29 条
[1]   NUCLEOTIDE-SEQUENCE OF A CDNA CLONE ENCODING HUMAN PREPROINSULIN [J].
BELL, GI ;
SWAIN, WF ;
PICTET, R ;
CORDELL, B ;
GOODMAN, HM ;
RUTTER, WJ .
NATURE, 1979, 282 (5738) :525-527
[2]   HUMAN FUR GENE ENCODES A YEAST KEX2-LIKE ENDOPROTEASE THAT CLEAVES PRO-BETA-NGF INVIVO [J].
BRESNAHAN, PA ;
LEDUC, R ;
THOMAS, L ;
THORNER, J ;
GIBSON, HL ;
BRAKE, AJ ;
BARR, PJ ;
THOMAS, G .
JOURNAL OF CELL BIOLOGY, 1990, 111 (06) :2851-2859
[3]   ISOLATION AND CHARACTERIZATION OF A CLONED RAT INSULIN GENE [J].
CORDELL, B ;
BELL, G ;
TISCHER, E ;
DENOTO, FM ;
ULLRICH, A ;
PICTET, R ;
RUTTER, WJ ;
GOODMAN, HM .
CELL, 1979, 18 (02) :533-543
[4]  
DAUGHERTY DF, 1991, AM J PHYSIOL, V260, pGM83
[5]  
DEBRUIJN MHL, 1985, P NATL ACAD SCI USA, V82, P708
[6]   INTRACELLULAR TARGETING AND STRUCTURAL CONSERVATION OF A PROHORMONE-PROCESSING ENDOPROTEASE [J].
FULLER, RS ;
BRAKE, AJ ;
THORNER, J .
SCIENCE, 1989, 246 (4929) :482-486
[7]   CONTINUOUS, CLONAL, INSULIN-SECRETING AND SOMATOSTATIN-SECRETING CELL-LINES ESTABLISHED FROM A TRANSPLANTABLE RAT ISLET CELL TUMOR [J].
GAZDAR, AF ;
CHICK, WL ;
OIE, HK ;
SIMS, HL ;
KING, DL ;
WEIR, GC ;
LAURIS, V .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (06) :3519-3523
[8]   PROCEDURE FOR MEASUREMENT OF DISTRIBUTION SPACES IN ISOLATED FAT CELLS [J].
GLIEMANN, J ;
GAMMELTOFT, S ;
VINTEN, J ;
OSTERLIND, K .
BIOCHIMICA ET BIOPHYSICA ACTA, 1972, 286 (01) :1-+
[9]   EXPRESSION OF SIMIAN VIRUS-40-RAT PREPROINSULIN RECOMBINANTS IN MONKEY KIDNEY-CELLS - USE OF PREPROINSULIN RNA PROCESSING SIGNALS [J].
GRUSS, P ;
KHOURY, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1981, 78 (01) :133-137
[10]  
HATSUZAWA K, 1990, J BIOL CHEM, V265, P22075