The membrane guanylyl cyclase, retinal guanylyl cyclase-1, is activated through its intracellular domain

被引:116
作者
Laura, RP
Dizhoor, AM
Hurley, JB
机构
[1] UNIV WASHINGTON, HOWARD HUGHES MED INST, SEATTLE, WA 98195 USA
[2] UNIV WASHINGTON, DEPT BIOCHEM, SEATTLE, WA 98195 USA
关键词
D O I
10.1074/jbc.271.20.11646
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
Retinal guanylyl cyclase-1 (RetGC-1) is a membrane guanylyl cyclase found in photoreceptor outer segments. It consists of an apparent extracellular domain (ECD) linked by a single transmembrane segment to an intracellular domain (ICD). Guanylyl cyclase activating protein-2 (GCAP-2) is a Ca2+-binding protein that activates RetGC-1 in a Ca2+-sensitive manner. To establish whether GCAP-2 stimulates RetGC-1 through the ECD or ICD, we made deletion mutants lacking either the ECD or both the ECD and transmembrane domains (TMD) of RetGC-1, Recombinant wild type RetGC-1 and both deletion mutants were expressed in HEK 293 cells, and their sensitivities to GCAP-2, Ca2+, and ATP were compared. Our data demonstrate that both deletion mutants are regulated similarly to wild type RetGC-1 with indistinguishable EC(50) values for Ca2+ and similar K-1/2 values for activation by GCAP-2, This shows that GCAP-2 functions through the ICD of RetGC-1 and that removal of the ECD and TMD do not significantly alter regulation by these factors. Our data also show that ATP potentiates stimulation of guanylyl cyclase activity by GCAP-2 and that neither the ECD nor the TMD of RetGC-1 participate in its regulation by ATP.
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页码:11646 / 11651
页数:6
相关论文
共 36 条
[1]   CA2+-DEPENDENT INTERACTION OF RECOVERIN WITH RHODOPSIN KINASE [J].
CHEN, CK ;
INGLESE, J ;
LEFKOWITZ, RJ ;
HURLEY, JB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (30) :18060-18066
[2]  
CHINKERS M, 1991, J BIOL CHEM, V266, P4088
[3]   CLONING, SEQUENCING, AND EXPRESSION OF A 24-KDA CA2+-BINDING PROTEIN ACTIVATING PHOTORECEPTOR GUANYLYL CYCLASE [J].
DIZHOOR, AM ;
OLSHEVSKAYA, EV ;
HENZEL, WJ ;
WONG, SC ;
STULTS, JT ;
ANKOUDINOVA, I ;
HURLEY, JB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (42) :25200-25206
[4]   RECOVERIN - A CALCIUM SENSITIVE ACTIVATOR OF RETINAL ROD GUANYLATE-CYCLASE [J].
DIZHOOR, AM ;
RAY, S ;
KUMAR, S ;
NIEMI, G ;
SPENCER, M ;
BROLLEY, D ;
WALSH, KA ;
PHILIPOV, PP ;
HURLEY, JB ;
STRYER, L .
SCIENCE, 1991, 251 (4996) :915-918
[5]   THE HUMAN PHOTORECEPTOR MEMBRANE GUANYLYL CYCLASE, RETGC, IS PRESENT IN OUTER SEGMENTS AND IS REGULATED BY CALCIUM AND A SOLUBLE ACTIVATOR [J].
DIZHOOR, AM ;
LOWE, DG ;
OLSHEVSKAYA, EV ;
LAURA, RP ;
HURLEY, JB .
NEURON, 1994, 12 (06) :1345-1352
[6]   CORE SEQUENCE OF ATP REGULATORY MODULE IN RECEPTOR GUANYLATE CYCLASES [J].
DUDA, T ;
GORACZNIAK, RM ;
SHARMA, RK .
FEBS LETTERS, 1993, 315 (02) :143-148
[7]   INDUCTION BY CYCLIC-GMP OF CATIONIC CONDUCTANCE IN PLASMA-MEMBRANE OF RETINAL ROD OUTER SEGMENT [J].
FESENKO, EE ;
KOLESNIKOV, SS ;
LYUBARSKY, AL .
NATURE, 1985, 313 (6000) :310-313
[8]  
GARBERS DL, 1994, J BIOL CHEM, V269, P30741
[9]  
GARBERS DL, 1992, CELL, V71, P1
[10]   STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF THE ROD OUTER SEGMENT MEMBRANE GUANYLATE-CYCLASE [J].
GORACZNIAK, RM ;
DUDA, T ;
SITARAMAYYA, A ;
SHARMA, RK .
BIOCHEMICAL JOURNAL, 1994, 302 :455-461