RGD Reference Report - Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis. - Rat Genome Database

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Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis.

Authors: Novakova, Z  Man, P  Novak, P  Hozak, P  Hodny, Z 
Citation: Novakova Z, etal., Electrophoresis. 2006 Apr;27(7):1277-87.
RGD ID: 10448962
Pubmed: PMID:16502463   (View Abstract at PubMed)
DOI: DOI:10.1002/elps.200500504   (Journal Full-text)

The nucleus is a highly structured organelle with distinct compartmentalization of specific functions. To understand the functions of these nuclear compartments, detailed description of protein complexes which form these structures is of crucial importance. We explored therefore the potential of blue native PAGE (BN-PAGE) method for the separation of nuclear protein complexes. We focused on (i) solubility and stability of nuclear complexes under conditions prerequisite for the separation by BN-PAGE, (ii) improved separation of native nuclear protein complexes using 2-D colorless native/blue native PAGE (CN-/BN-PAGE), and (iii) mass spectrometric analysis of protein complexes which were isolated directly from native 1-D or from 2-D CN/BN-PAGE gels. The suitability of BN-PAGE for nuclear proteomic research is demonstrated by the successful separation of polymerase I and polymerase II complexes, and by mass spectrometric determination of U1 small nuclear ribonucleoprotein particle composition. Moreover, practical advice for sample preparation is provided.



Gene Ontology Annotations    Click to see Annotation Detail View

Cellular Component

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Snrnp70RatU1 snRNP  IDA  RGD 
SnrpaRatU1 snRNP  IDA  RGD 
SnrpcRatU1 snRNP  IDA  RGD 
Snrpd1RatU1 snRNP  IDA  RGD 
Snrpd2RatU1 snRNP  IDA  RGD 
Snrpd3RatU1 snRNP  IDA  RGD 
SnrpeRatU1 snRNP  IDA  RGD 
SnrpgRatU1 snRNP  IDA  RGD 

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Snrnp70RatU1 snRNP binding  IDA  RGD 
SnrpaRatU1 snRNP binding  IDA  RGD 
SnrpcRatU1 snRNP binding  IDA  RGD 
Snrpd1RatU1 snRNP binding  IDA  RGD 
Snrpd2RatU1 snRNP binding  IDA  RGD 
Snrpd3RatU1 snRNP binding  IDA  RGD 
SnrpeRatU1 snRNP binding  IDA  RGD 
SnrpgRatU1 snRNP binding  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Snrnp70  (small nuclear ribonucleoprotein U1 subunit 70)
Snrpa  (small nuclear ribonucleoprotein polypeptide A)
Snrpc  (small nuclear ribonucleoprotein polypeptide C)
Snrpd1  (small nuclear ribonucleoprotein D1 polypeptide)
Snrpd2  (small nuclear ribonucleoprotein D2 polypeptide)
Snrpd3  (small nuclear ribonucleoprotein D3 polypeptide)
Snrpe  (small nuclear ribonucleoprotein polypeptide E)
Snrpg  (small nuclear ribonucleoprotein polypeptide G)


Additional Information