The purposes for which the fractionation of proteins are
carried out are quite varied and manyfold. However, one of the
more important reasons is that of determining the nature and the
extent of autolysis or proteolysis on the bovine muscle proteins
during post mortem aging. Hence, the development of a procedure
for the adequate fractionation of muscle proteins would greatly stimulate
the interest and research progress in this difficult field of study.
The research reported herein pertains to a study of the
fractionation of fresh bovine muscle proteins by ion exchange chromatography.
A KCl-phosphate buffer, pH 7.5 and an ionic strength
of 0.55, was used to extract the proteins from the muscle. This extract
was then diluted to specific ionic strengths in order to separate
the gross fractions (actomyosin, myosin, sarcoplasmic + actin) from the total KCl-phosphate soluble proteins. The major protein
fractions were then separated by diethylaminoethyl-cellulose (DEAE-cellulose)
ion exchange chromatographic procedure. A non-linear
gradient elution schedule was used throughout the chromatographic
procedure. The disc electrophoresis technique was used to determine
the homogeneity of the various protein fractions and sub-fractions.
The total KCl-phosphate soluble proteins were fractionated
into 9-12 fractions by DEAE-cellulose ion exchange chromatography.
The number of fractions differed from one sample to another. The
disc electrophoresis results paralleled those of the chromatographic
procedure. Some of the fractions appeared to be homogeneous while
others were not.
The KCl-phosphate soluble proteins minus actomyosin were
fractionated by column chromatography into 9-10 different protein
components. The disc electrophoresis results also indicated that
this fraction contained 9-10 components.
The sarcoplasmic + actin proteins were fractionated into
six fractions by the chromatographic procedure. These fractions
appeared to be electrophoretically homogeneous.
Although some success was attained in the separation of
the actomyosin proteins, the myosin fraction was quite resistant to
chromatographic separation.
In spite of the ineffectiveness of the DEAE-cellulose column
chromatographic technique to fractionate the myosin and actomyosin
proteins, the procedure appears to have some value for studying
the other protein fractions during autolysis. Moreover, further
work on the adoption of this procedure might provide the necessary
information to perfect the technique for muscle protein fractionation. / Graduation date: 1963
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/27384 |
| Date | 15 May 1963 |
| Creators | Mohasseb, Zeinab Shehata |
| Contributors | Anglemier, Allen F. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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