Premium
Further Physicochemical Characterization of the Novel Human Brain Protein h 3
Author(s) -
Bollengier F.,
Beeckmans S.,
Mahler A.,
Kanarek L.
Publication year - 1989
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1989.tb01856.x
Subject(s) - chemistry , circular dichroism , sodium dodecyl sulfate , polyacrylamide gel electrophoresis , titration , gel electrophoresis , polyacrylamide , disulfide bond , absorption (acoustics) , reagent , crystallography , biochemistry , enzyme , inorganic chemistry , polymer chemistry , organic chemistry , physics , acoustics
Recently we reported the isolation and partial biochemical characterization of the novel polypeptide h 3 from human brain and liver. In this report, the physicochemical characterization is further established by the use of several analytical methods. The following results were obtained: the ultraviolet absorption spectrum is not influenced by pH, and the circular dichroism (CD) spectrum reveals that this protein has no a‐helices, whereas ± 25% of the polypeptide chain is found to be folded as a β ‐pleated sheet structure. Neither the conformation of h 3 as assessed by CD nor the titration kinetics of sulfhydryl groups with Ellman's reagent are affected by the presence of the ions K + , Na + , Ca 2+ , and Mg 2+ . Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) in a β ‐mercaptoethanol gradient and Cleveland sequential SDS‐PAGE showed that the frequent formation of h 3 polymers and doublets, as observed earlier, is almost exclusively due to disulfide bonding.