Stabilization of the nuclear matrix by disulfide bridges: identification of matrix polypeptides that form disulfides.
The molecular structure of the nuclear matrix is still poorly understood. We have tried to assess which proteins are important structural elements by examining the process of stabilization of the nuclear matrix by sodium tetrathionate. Sodium tetrathionate stabilizes the nuclear matrix by oxidizing sulfhydryl groups to disulfides. We show that tetrathionate-stabilized matrices are disassembled in buffers containing SDS, indicating that the stabilized nuclear matrix is not a continuous network of cross-linked proteins. Using monobromobimane, a thiol-specific fluorescent reagent, we show that many protein thiols in the stabilized matrix are oxidized. By chromatography on activated thiol-Sepharose we estimated that about 50% of the matrix proteins had oxidized sulfhydryl groups. The protein composition of the material bound to activated thiol-Sepharose was similar to that of the not-bound material. A few proteins are highly enriched in the fraction that was bound to the column. This indicates that many matrix protein species are partially oxidized and that some proteins are completely oxidized. The oxidized protein thiols are found in relatively large complexes as determined by SDS gel-electrophoresis under nonreducing conditions. These results are interpreted in terms of protein-protein interactions in the matrix. The possible role of thiols and disulfides in the in vivo organization of the nucleus is discussed.
Stuurman
N
N
E. C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
Floore
A
A
Colen
A
A
de Jong
L
L
van Driel
R
R
D000818:Animals; D003432:Cross-Linking Reagents; D004220:Disulfides; D066298:In Vitro Techniques; D034882:Lamins; D046911:Macromolecular Substances; D008970:Molecular Weight; D015530:Nuclear Matrix / Q000737:chemistry / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000737:chemistry*; D010084:Oxidation-Reduction; D051381:Rats; D013776:Tetrathionic Acid / Q000737:chemistry
D003432:Cross-Linking Reagents; D004220:Disulfides; D034882:Lamins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D013776:Tetrathionic Acid