NDLI: Protein phosphorylation corrects the folding defect of the neuroblastoma (S120G) mutant of human nucleoside diphosphate kinase A/Nm23-H1

Content Provider	PubMed Central
Author	Mocan, Iulia Florian, Georgescauld Gonin, Philippe Thoraval, Didier Cervoni, Laura Anna, Giartosio Dabernat-arnaud, Sandrine Crouzet, Marc Lacombe, Marie-lise Lascu, Ioan
Copyright Year	2007
Abstract	Human nucleoside diphosphate (NDP) kinase A is a ‘house-keeping’ enzyme essential for the synthesis of nonadenine nucleoside (and deoxynucleoside) 5′-triphosphate. It is involved in complex cellular regulatory functions including the control of metastatic tumour dissemination. The mutation S120G has been identified in high-grade neuroblastomas. We have shown previously that this mutant has a folding defect: the urea-denatured protein could not refold in vitro. A molten globule folding intermediate accumulated, whereas the wild-type protein folded and associated into active hexamers. In the present study, we report that autophosphorylation of the protein corrected the folding defect. The phosphorylated S120G mutant NDP kinase, either autophosphorylated with ATP as donor, or chemically prosphorylated by phosphoramidate, refolded and associated quickly with high yield. Nucleotide binding had only a small effect. ADP and the non-hydrolysable ATP analogue 5′-adenyly-limido-diphosphate did not promote refolding. ATP-promoted refolding was strongly inhibited by ADP, indicating protein dephosphorylation. Our findings explain why the mutant enzyme is produced in mammalian cells and in Escherichia coli in a soluble form and is active, despite the folding defect of the S120G mutant observed in vitro. We generated an inactive mutant kinase by replacing the essential active-site histidine residue at position 118 with an asparagine residue, which abrogates the autophosphorylation. The double mutant H118N/S120G was expressed in inclusion bodies in E. coli. Its renaturation stops at a folding intermediate and cannot be reactivated by ATP in vitro. The transfection of cells with this double mutant might be a good model to study the cellular effects of folding intermediates.
Related Links	http://dx.doi.org/10.1042/bj20061141
Ending Page	156
Page Count	8
Starting Page	149
File Format	PDF
ISSN	02646021
e-ISSN	14708728
Journal	Biochemical Journal
Issue Number	Pt 1
Volume Number	403
Language	English
Publisher	Portland Press Ltd.
Publisher Date	2007-04-01
Access Restriction	Open
Rights Holder	Portland Press Ltd.
Subject Keyword	Cell Biology Biochemistry Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Molecular Biology Biochemistry

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Protein phosphorylation corrects the folding defect of the neuroblastoma (S120G) mutant of the human nucleoside diphosphate kinase A/Nm23-H1

Crystal Structures of S120G Mutant and Wild Type of Human Nucleoside Diphosphate Kinase A in Complex with ADP

Nm23-H1/NDP kinase folding intermediates and cancer: a hypothesis

A Point Mutation of Human Nucleoside Diphosphate Kinase A Found in Aggressive Neuroblastoma Affects Protein Folding

The Nucleoside Diphosphate Kinase Nm23-H4 Acts as a Mitochondrial Switch♦

Nucleoside diphosphate kinase/Nm23 and Epstein–Barr virus

Glyceraldehyde-3-phosphate Dehydrogenase and Nm23-H1/Nucleoside Diphosphate Kinase A TWO OLD ENZYMES COMBINE FOR THE NOVEL Nm23 PROTEIN PHOSPHOTRANSFERASE FUNCTION

Role of Binding and Nucleoside Diphosphate Kinase A in the Regulation of the Cystic Fibrosis Transmembrane Conductance Regulator by AMP-activated Protein Kinase

Regulatory roles for nm23/nucleoside diphosphate kinase-like enzymes in insulin secretion from the pancreatic islet beta cell

Protein phosphorylation corrects the folding defect of the neuroblastoma (S120G) mutant of human nucleoside diphosphate kinase A/Nm23-H1

Similar Documents

Protein phosphorylation corrects the folding defect of the neuroblastoma (S120G) mutant of the human nucleoside diphosphate kinase A/Nm23-H1

Crystal Structures of S120G Mutant and Wild Type of Human Nucleoside Diphosphate Kinase A in Complex with ADP

Nm23-H1/NDP kinase folding intermediates and cancer: a hypothesis

A Point Mutation of Human Nucleoside Diphosphate Kinase A Found in Aggressive Neuroblastoma Affects Protein Folding

The Nucleoside Diphosphate Kinase Nm23-H4 Acts as a Mitochondrial Switch♦

Nucleoside diphosphate kinase/Nm23 and Epstein–Barr virus

Glyceraldehyde-3-phosphate Dehydrogenase and Nm23-H1/Nucleoside Diphosphate Kinase A TWO OLD ENZYMES COMBINE FOR THE NOVEL Nm23 PROTEIN PHOSPHOTRANSFERASE FUNCTION

Role of Binding and Nucleoside Diphosphate Kinase A in the Regulation of the Cystic Fibrosis Transmembrane Conductance Regulator by AMP-activated Protein Kinase

Regulatory roles for nm23/nucleoside diphosphate kinase-like enzymes in insulin secretion from the pancreatic islet beta cell

Protein phosphorylation corrects the folding defect of the neuroblastoma (S120G) mutant of human nucleoside diphosphate kinase A/Nm23-H1