Post-translational processing of Drosophila nucleoside diphosphate kinase.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/109159
- author
- Stenberg, Leisa LU ; Stenflo, Johan LU ; Holmgren, Paul LU and Brown, Mark A. LU
- organization
- publishing date
- 2002
- type
- Contribution to journal
- publication status
- published
- subject
- 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
- 2016-04-01 15:41:18
- date last changed
- 2022-01-28 06:33:05
@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}},
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}},
language = {{eng}},
number = {{3}},
pages = {{689--694}},
publisher = {{Elsevier}},
series = {{Biochemical and Biophysical Research Communications}},
title = {{Post-translational processing of Drosophila nucleoside diphosphate kinase.}},
url = {{http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12099695&dopt=Abstract}},
volume = {{295}},
year = {{2002}},
}