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Similarity between nuclear matrix proteins of various cells revealed by an improved isolation method
Author(s) -
Gerner Christopher,
Holzmann Klaus,
Grimm Rudolf,
Sauermann Georg
Publication year - 1998
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19981201)71:3<363::aid-jcb5>3.0.co;2-w
Subject(s) - nuclear matrix , nuclear protein , nuclear lamina , ribonucleoprotein , lamin , biology , cell nucleus , microbiology and biotechnology , heterogeneous nuclear ribonucleoprotein , chemistry , biochemistry , rna , dna , nucleus , chromatin , gene , transcription factor
Comparative analysis of nuclear matrix proteins by two‐dimensional electrophoresis may be greatly impaired by copurifying cytoskeletal proteins. The present data show that the bulk of adhering cytofilaments may mechanically be removed by shearing of nuclei pretreated with vanadyl ribonucleoside complexes. Potential mechanisms of action not based on ribonuclease inhibition are discussed. To individually preserve the integrity of nuclear structures, we developed protocols for the preparation of nuclear matrices from three categories of cells, namely leukocytes, cultured cells, and tissue cells. As exemplified with material from human lymphocytes, cultured amniotic cells, and liver tissue cells, the resulting patterns of nuclear matrix proteins appeared quite similar. Approximately 300 spots were shared among the cell types. Forty‐nine of these were identified, 21 comprising heterogeneous nuclear ribonucleoproteins. Heterogeneous nuclear ribonucleoproteins L and nuclear lamin B2 isoforms were identified by amino acid sequencing and mass spectrometry. However, individually expressed proteins, such as the proliferating cell nuclear antigen, also pertained following application of the protocols. Thus, enhanced resolution and comparability of proteins improve systematic analyses of nuclear matrix proteins from various cellular sources. J. Cell. Biochem. 71:363–374, 1998. © 1998 Wiley‐Liss, Inc.