+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Functional cloning of a nucleoside diphosphate kinase from Dictyostelium discoideum

, : Functional cloning of a nucleoside diphosphate kinase from Dictyostelium discoideum. Journal of Biological Chemistry 265(17): 10012-8

A .lambda.gt11 cDNA library from Dictyostelium discoideum was screened by direct labeling of filter replicas with [35S]guanosine 5'-O-(thiotriphosphate) (GTP.gamma.S). A positive clone was obtained and used as probe to isolate additional clones from which a complete cDNA sequence was determined. The cDNA hybridizes to a single copy gene that is expressed as a 0.6-kilobase mRNA in vegetatively growing amoeba. The open reading frame encodes a protein of 155 amino acids (calculated Mr 16,775), devoid of cysteine residues. The protein contains most of the short consensus motifs characteristic of the catalytic domain of protein kinases although the overall homology with this class of enzymes is not greater than 20%. Its size and amino acid composition indicated that it could be the monomer of a nucleoside diphosphate (NDP)-kinase, an enzyme which catalyzes the phosphate transfer from triphospho- to diphosphonucleotides. Indeed, specific NDP kinase activity was found in extracts of bacteria transformed with a plasmid expressing the protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the extracts incubated in the presence of [35S]GTP.gamma.S revealed a single 35S-labeled band of size corresponding to the protein, which likely represents the stable thiophosphorylated reaction intermediate characteristic for the ping-pong reaction mechanism of NDP kinases. The formation of this labeled intermediate probably allowed the detection of the enzyme on the filters during the screening procedure. Although NDP kinases from a great variety of sources have been characterized, the primary structure of the D. discoideum NDP kinase is the first reported for an enzyme of eukaryotic origin.


PMID: 2161830

Other references

Parkhomenko, O.V., 2012: A giant prostatic adenoma without disturbance of patient's quality of life. Urologiia: 77-77

Raiha, N.C.R., 1976: Milk protein, quality and quantity, in the feeding of low birth weight infants. Studies indicate that when low birth wt. infants are fed a diet relatively high in casein-predominant milk protein, they have high plasma concn. of the essential amino acids and tyrosine for up to 6 wk of age. In addition, they show high urea N an...

Birrane, G.; Soni, A.; Ladias, J.A.A., 2009: Structural basis for DNA recognition by the human PAX3 homeodomain. The transcription regulatory protein PAX3 binds to cognate DNA sequences through two DNA-binding domains, a paired domain and a homeodomain, and has important functions during neurogenesis and myogenesis. In humans, mutations in the PAX3 gene caus...

Auguste, C.; Paris, J., 1955: Study of the jejunal fluid in a case of sprue nostras. Archives des Maladies de L'appareil Digestif et des Maladies de la Nutrition 44(11): 1200-1207

Uppal, P.K.; Yadav, M.P., 1990: Equine infectious anaemia: experimental transmission in donkeys. Results indicate that equine infectious anaemia can be transmitted from symptomatic or asymptomatic seropositive horses to indigenous Indian donkeys, by subcutaneous or intravenous inoculation of blood, spleen or lymph node suspensions, and from d...

Atkinson, J.; Braddick, O., 1983: Assessment of visual acuity in infancy and early childhood. The forced-choice preferential looking method (FPL) shows the development of acuity during the first year of life, and is applicable to clinical assessment. A tracking test using a narrow strip of grating yields a more sensitive measure for the la...

Melega, W.P.; Lacan, G.; Way, B.M.; Jorgensen, M.J.; Morton, G.; Pham, J.; Fairbanks, L.A., 2003: Alterations in brain and behavior in the vervet monkey after chronic methamphetamine. A chronic methamphetamine (METH) protocol was administered over a 6 month period to socially housed adult vervet monkeys (Cercopithecus aethiops, n = 5) to simulate aspects of a prevalent human METH abuse pattern. METH exposure included escalating...

Ward, J.M.; Rice, J.M., 1982: Naturally occurring and chemically induced brain tumors of rats and mice in carcinogenesis bioassays. Annals of the New York Academy of Sciences 381: 304-319

Collins, K.K.; Schaffer, M.S., 2011: Use of cryoablation for treatment of tachyarrhythmias in 2010: survey of current practices of pediatric electrophysiologists. Cryoablation for arrhythmia substrates in pediatrics has been available since 2003. The purpose of this study was to evaluate the current approach of pediatric electrophysiologists to the use of cryoablation in the current era. We sent an Internet...

Melkote, S.R.; Muppidi, S.R.; Goswami, D., 2000: Social and economic factors in an integrated behavioral and societal approach to communications in HIV/AIDS. Health communication scholars have tried to understand how individuals process information and have identified the factors that contribute to appropriate behavior change. Some of these theorists have, implicitly or explicitly, assumed that if indi...