Catalytic mechanism of the adenylyl and guanylyl cyclases: Modeling and mutational analysis

被引:231
作者
Liu, Y
Ruoho, AE
Rao, VD
Hurley, JH
机构
[1] NIDDKD,MOL BIOL LAB,NIH,BETHESDA,MD 20892
[2] UNIV WISCONSIN,SCH MED,DEPT PHARMACOL,MADISON,WI 53706
关键词
D O I
10.1073/pnas.94.25.13414
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The adenylyl and guanylyl cyclases catslyze the formation of 3 ',5 ' cyclic adenosine or guanosine monophosphate from the corresponding nucleoside 5 '-triphosphate. The guanylyl cyclases, the mammalian adenylyl cyclases, and their microbial homologues function as pairs of homologous catalytic domains. The crystal structure of the rat type II adenylyl cyclase C-2 catalytic domain was used to model by homology a mammalian adenylyl cyclase C-1-C-2 domain pair, a homodimeric soluble cyclase of dictyostelium discoideum, a heterodimeric soluble guanylyl cyclase, and a homodimeric membrane guanyly cyclase. Mg(2+)ATP or Mg(2+)GTP were docked into the active sites based on known stereochemical constraints on their conformation. The models are consistent with the activities of seven active-site mutants. Asp-310 and Glu-432 of type 1 adenylyl cyclase coordinate a Mg(2+)ion. The D310S and D310A mutants have lO-fold reduced V-max and altered [Mg2+] dependence. The NTP purine moieties bind in mostly hydrophobic pockets. Specificity is conferred by a Lys and an Asp in adenylyl cyclase, and a Glu, an Arg, and a Cys in guanyly cyclase. The models predict that an Asp from one domain is a general base in the reaction, and that the transition state is stabilized by a conserved Asn-Arg pair on the other domain.
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页码:13414 / 13419
页数:6
相关论文
共 53 条
[1]  
Artymiuk PJ, 1997, NATURE, V388, P33, DOI 10.1038/40310
[2]   CRYSTAL-STRUCTURES OF THE KLENOW FRAGMENT OF DNA-POLYMERASE-I COMPLEXED WITH DEOXYNUCLEOSIDE TRIPHOSPHATE AND PYROPHOSPHATE [J].
BEESE, LS ;
FRIEDMAN, JM ;
STEITZ, TA .
BIOCHEMISTRY, 1993, 32 (51) :14095-14101
[3]   FROM ADENYLATE-CYCLASE TO GUANYLATE-CYCLASE - MUTATIONAL ANALYSIS OF A CHANGE IN SUBSTRATE-SPECIFICITY [J].
BEUVE, A ;
DANCHIN, A .
JOURNAL OF MOLECULAR BIOLOGY, 1992, 225 (04) :933-938
[4]  
BRUNGER AT, 1996, XPLOR
[5]   A polymerase I palm in adenylyl cyclase? Reply [J].
Bryant, SH ;
Madej, T ;
Janin, J ;
Liu, Y ;
Ruoho, AE ;
Zhang, GY ;
Hurley, JH .
NATURE, 1997, 388 (6637) :34-34
[6]   RIBBON MODELS OF MACROMOLECULES [J].
CARSON, M .
JOURNAL OF MOLECULAR GRAPHICS, 1987, 5 (02) :103-&
[7]   A REGION OF ADENYLYL-CYCLASE-2 CRITICAL FOR REGULATION BY G-PROTEIN BETA-GAMMA-SUBUNITS [J].
CHEN, JQ ;
DEVIVO, M ;
DINGUS, J ;
HARRY, A ;
LI, JR ;
SUI, JL ;
CARTY, DJ ;
BLANK, JL ;
EXTON, JH ;
STOFFEL, RH ;
INGLESE, J ;
LEFKOWITZ, RJ ;
LOGOTHETIS, DE ;
HILDEBRANDT, JD ;
IYENGAR, R .
SCIENCE, 1995, 268 (5214) :1166-1169
[8]   STRUCTURES OF ACTIVE CONFORMATIONS OF G(I-ALPHA-1) AND THE MECHANISM OF GTP HYDROLYSIS [J].
COLEMAN, DE ;
BERGHUIS, AM ;
LEE, E ;
LINDER, ME ;
GILMAN, AG ;
SPRANG, SR .
SCIENCE, 1994, 265 (5177) :1405-1412
[9]   Inhibition of adenylyl cyclase by a family of newly synthesized adenine nucleoside 3'-polyphosphates [J].
Desaubry, L ;
Shoshani, N ;
Johnson, RA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (24) :14028-14034
[10]   Interactions of forskolin and ATP with the cytosolic domains of mammalian adenylyl cyclase [J].
Dessauer, CW ;
Scully, TT ;
Gilman, AG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (35) :22272-22277