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Bifunctional reagents and protein structure determination. The reaction of phenolic disulfonyl chlorides with lysozyme
Author(s) -
Herzig David J.,
Rees Allan W.,
Day Richard A.
Publication year - 1964
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1964.360020406
Subject(s) - lysozyme , chemistry , bifunctional , chloride , phenol , hydrolysis , reagent , covalent bond , lysine , chromatography , trypsin , aqueous solution , organic chemistry , enzyme , catalysis , amino acid , biochemistry
It was established that phenol‐2,4‐disulfonyl chloride and α‐naphthol‐2, 4‐disulfonyl chloride introduce new intramolecular or tertiary bonds into lysozyme. The phenolic hydroxyl group seems to make the cross linking reagents soluble in aqueous media; it was established that a sulfonylating; species does dissolve prior to hydrolysis but that the chlorosulfonyl groups are hydrolyzed within seconds of dissolution. New covalently linked residues were shown to be introduced by different, spectra. Peptide maps of the lysozyme treated with phenol‐2,4‐disulfonyl chloride. peroxidized, and digested with trypsin showed definite deletions and additions when compared with the control. Partial analysis of the peptides showed that cross linking between ε‐amino groups of lysine residues had occurred. A certain degree of stabilization of lysozyme to inactivation by certain inactivating conditions was noted after treatment with the two reagents. Ultra‐centrifugal analysis of the lysozyme treated with phenol‐2,4‐disulfonyl chloride showed that there was no detectable fraction having a higher molecular weight than native lysozyme. Tentative assignments of positions of cross linking were made.