Resistance of chromatin superstructure to tryptic digestion modulated by conjugated polyacrylamide

FEBS Letters

Skinner, J.D.¹  Burgoyne, L.A.¹  Saccone, G.T.P.¹ 

[1] School of Biological Sciences, Flinders University of South Australia, Bedford Park, SA 5042, Australia

关键词: Chromatin;    Nucleoprotein;    Histone;    Trypsin;    Erythrocyte;    Liver nuclei;    PA—trypsin;    soluble polyacrylamide—trypsin conjugate;    trypsin—TPCK;    L-(tosylamido-2-phenyl)ethyl chloromethyl-treated trypsin;    TEMED;    N;    NN′;    N-tetramethylethylbenediamine;    Hepes;    N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid;    M r;    relative molecular mass;   

DOI  :  10.1016/0014-5793(83)81126-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Soluble polyacrylamide conjugates have been used to modulate tryptic digestion of chromatin. Digestion of histones H₁/H₅ and H₃ are mutually dependent and relatively independent of that of the core histones which are not significantly digested until 50–60% of H₃ is degraded. H₁/H₅ and H₃ are most exposed, H₃ behaves as a ‘non-core’ histone, its destruction appears to be the critical factor in the collapse of the chromatin superstructure during tryptic digestions. The digestion kinetics are explained by proposing that the initial sites of attack are in cavities much larger than the diameter of trypsin (4–5 nm). Procedures for the preparation of soluble polyacrylamide and its conjugation to trypsin are described.

【 授权许可】

Unknown