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Post-translational processing of Drosophila nucleoside diphosphate kinase.

Stenberg, Leisa LU ; Stenflo, Johan LU ; Holmgren, Paul LU and Brown, Mark A. LU (2002) In Biochemical and Biophysical Research Communications 295(3). p.689-694
Abstract
Nucleoside diphosphate kinase (NDPK) was purified from Drosophila melanogaster by a combination of anion-exchange, hydroxyapatite, and reversed-phase chromatography. The identity of the purified enzyme was confirmed by sequencing internal peptides (the N-terminus appeared to be blocked). Post-translational modifications were investigated by using protein chemical and mass spectrometric methods. Analysis by nanoelectrospray ionization-mass spectrometry revealed that the mass of the enzyme was considerably smaller than that predicted from its amino acid sequence. Although its open-reading frame predicts a 153-residue polypeptide, the mature enzyme was found to comprise 152 amino acids, being modified by proteolytic removal of the initiator... (More)
Nucleoside diphosphate kinase (NDPK) was purified from Drosophila melanogaster by a combination of anion-exchange, hydroxyapatite, and reversed-phase chromatography. The identity of the purified enzyme was confirmed by sequencing internal peptides (the N-terminus appeared to be blocked). Post-translational modifications were investigated by using protein chemical and mass spectrometric methods. Analysis by nanoelectrospray ionization-mass spectrometry revealed that the mass of the enzyme was considerably smaller than that predicted from its amino acid sequence. Although its open-reading frame predicts a 153-residue polypeptide, the mature enzyme was found to comprise 152 amino acids, being modified by proteolytic removal of the initiator Met and N-acetylation of Ala2. This explains why the observed pI of the Drosophila enzyme is more acidic than that predicted from its amino acid sequence. No additional post-translational modifications such as glycosylation or O-phosphorylation, which have been identified on homologous NDPKs from other organisms, were detected on the Drosophila enzyme. (Less)
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keywords
Drosophila melanogaster : enzymology, Durapatite : pharmacology, Glycosylation, Nucleoside-Diphosphate Kinase : chemistry, Nucleoside-Diphosphate Kinase : isolation & purification, Open Reading Frames, Peptides : chemistry, Phosphorylation, Protein Processing, Spectrum Analysis, Post-Translational, Mass, Ion Exchange, Chromatography, Biocompatible Materials : pharmacology, Animal
in
Biochemical and Biophysical Research Communications
volume
295
issue
3
pages
689 - 694
publisher
Elsevier
external identifiers
  • wos:000176908000020
  • pmid:12099695
  • scopus:0036310978
ISSN
1090-2104
language
English
LU publication?
yes
id
8cdc2ec3-f64d-4c7f-bf8c-ae3ea9c65163 (old id 109159)
alternative location
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12099695&dopt=Abstract
date added to LUP
2007-07-27 09:31:38
date last changed
2017-06-04 04:17:10
@article{8cdc2ec3-f64d-4c7f-bf8c-ae3ea9c65163,
  abstract     = {Nucleoside diphosphate kinase (NDPK) was purified from Drosophila melanogaster by a combination of anion-exchange, hydroxyapatite, and reversed-phase chromatography. The identity of the purified enzyme was confirmed by sequencing internal peptides (the N-terminus appeared to be blocked). Post-translational modifications were investigated by using protein chemical and mass spectrometric methods. Analysis by nanoelectrospray ionization-mass spectrometry revealed that the mass of the enzyme was considerably smaller than that predicted from its amino acid sequence. Although its open-reading frame predicts a 153-residue polypeptide, the mature enzyme was found to comprise 152 amino acids, being modified by proteolytic removal of the initiator Met and N-acetylation of Ala2. This explains why the observed pI of the Drosophila enzyme is more acidic than that predicted from its amino acid sequence. No additional post-translational modifications such as glycosylation or O-phosphorylation, which have been identified on homologous NDPKs from other organisms, were detected on the Drosophila enzyme.},
  author       = {Stenberg, Leisa and Stenflo, Johan and Holmgren, Paul and Brown, Mark A.},
  issn         = {1090-2104},
  keyword      = {Drosophila melanogaster : enzymology,Durapatite : pharmacology,Glycosylation,Nucleoside-Diphosphate Kinase : chemistry,Nucleoside-Diphosphate Kinase : isolation & purification,Open Reading Frames,Peptides : chemistry,Phosphorylation,Protein Processing,Spectrum Analysis,Post-Translational,Mass,Ion Exchange,Chromatography,Biocompatible Materials : pharmacology,Animal},
  language     = {eng},
  number       = {3},
  pages        = {689--694},
  publisher    = {Elsevier},
  series       = {Biochemical and Biophysical Research Communications},
  title        = {Post-translational processing of Drosophila nucleoside diphosphate kinase.},
  volume       = {295},
  year         = {2002},
}