Trypanosoma cruzi arginine kinase characterization and cloning -: A novel energetic pathway in protozoan parasites

被引:143
作者
Pereira, CA
Alonso, GD
Paveto, MC
Iribarren, A
Cabanas, ML
Torres, HN
Flawiá, MM
机构
[1] Univ Buenos Aires, Consejo Nacl Invest Cient & Tecn, Inst Invest Ingn Genet & Biol Mol, Buenos Aires, DF, Argentina
[2] Univ Buenos Aires, Fac Ciencias Exactas & Nat, Buenos Aires, DF, Argentina
关键词
D O I
10.1074/jbc.275.2.1495
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
This work contains the first description of a guanidino kinase in a flagellar unicellular parasite. The enzyme phosphorylates L-arginine and was characterized in preparations from Trypanosoma cruzi, the ethiological agent of Chagas' disease. The activity requires ATP and a divalent cation. Under standard assay conditions (1 mM L-arginine), the presence of B-fold higher concentrations of canavanine or histidine produced a greater than 50% enzyme inhibition, The base sequence of this enzyme revealed an open reading frame of 357 amino acids and a molecular weight of 40,201. The amino acid sequence shows all of the characteristic consensus blocks of the ATP:guanidino phosphotransferase family and a putative "actinin-type" actin-binding domain. The highest amino acid identities of the T. cruzi sequence, about 70%, were with arginine kinases from Arthropoda. Southern and chromosome blots revealed that the kinase is encoded by a single-copy gene. Moreover, Northern blot analysis showed an mRNA subpopulation of about 2.0 kilobases, and Western blotting of T. cruzi-soluble polypeptides revealed a 40-kDa band. The finding in the parasite of a phosphagen and its biosynthetic pathway, which are totally different from those in mammalian host tissues, points out this arginine kinase as a possible chemotherapy target for Chagas' disease.
引用
收藏
页码:1495 / 1501
页数:7
相关论文
共 22 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]  
CAMARGO EP, 1978, EXP PARASITOL, V46, P141, DOI 10.1016/0014-4894(78)90125-X
[3]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[4]   TRYPANOSOMA-CRUZI EXHIBITS INTER-STRAIN AND INTRA-STRAIN HETEROGENEITY IN MOLECULAR KARYOTYPE AND CHROMOSOMAL GENE LOCATION [J].
ENGMAN, DM ;
REDDY, LV ;
DONELSON, JE ;
KIRCHHOFF, LV .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1987, 22 (2-3) :115-123
[5]   Purification, characterization, and hydrodynamic properties of arginine kinase from Gulf shrimp (Penaeus aztecus) [J].
France, RM ;
Sellers, DS ;
Grossman, SH .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1997, 345 (01) :73-78
[6]   New features of the blocks database servers [J].
Henikoff, JG ;
Henikoff, S ;
Pietrokovski, S .
NUCLEIC ACIDS RESEARCH, 1999, 27 (01) :226-228
[7]   CLUSTAL - A PACKAGE FOR PERFORMING MULTIPLE SEQUENCE ALIGNMENT ON A MICROCOMPUTER [J].
HIGGINS, DG ;
SHARP, PM .
GENE, 1988, 73 (01) :237-244
[8]  
Huennekens FM, 1960, COMP BIOCHEM, p[107, 1]
[9]   THE IDENTIFICATION AND CHARACTERIZATION OF AN ACTIN-BINDING SITE IN ALPHA-ACTININ BY MUTAGENESIS [J].
KUHLMAN, PA ;
HEMMINGS, L ;
CRITCHLEY, DR .
FEBS LETTERS, 1992, 304 (2-3) :201-206
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+