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Heterogeneity Determination and Purification of Commercial Hen Egg-White LysozymeHen egg-white lysozyme (HEWL) is widely used as a model protein, although its purity has not been adequately characterized by modern biochemical techniques. We have identified and quantified the protein heterogeneities in three commercial HEWL preparations by sodium dodecyl sulfate polyacrylamide gel electrophoresis with enhanced silver staining, reversed-phase fast protein liquid chromatography (FPLC) and immunoblotting with comparison to authentic protein standards. Depending on the source, the contaminating proteins totalled 1-6%(w/w) and consisted of ovotransferrin, ovalbumin, HEWL dimers, and polypeptides with approximate M(sub r) of 39 and 18 kDa. Furthermore, we have obtained gram quantities of electrophoretically homogeneous [> 99.9%(w/w)] HEWL by single-step semi-preparative scale cation-exchange FPLC with a yield of about 50%. Parallel studies of crystal growth kinetics, salt repartitioning and crystal perfection with this highly purified material showed fourfold increases in the growth-step velocities and significant enhancement in the structural homogeneity of HEWL crystals.
Document ID
19990046762
Acquisition Source
Marshall Space Flight Center
Document Type
Reprint (Version printed in journal)
Authors
Thomas, B. R.
(Alabama Univ. Huntsville, AL United States)
Vekilov, P. G.
(Alabama Univ. Huntsville, AL United States)
Rosenberger, F.
(Alabama Univ. Huntsville, AL United States)
Date Acquired
August 19, 2013
Publication Date
January 1, 1998
Publication Information
Publication: Acta Crystallographica Section D
Publisher: International Union of Crystallography
Volume: D52
ISSN: 0907-4449
Subject Category
Chemistry And Materials (General)
Funding Number(s)
CONTRACT_GRANT: NAG8-950
CONTRACT_GRANT: NAG8-1161
Distribution Limits
Public
Copyright
Other

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