Date of Award

Summer 1984

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Chemistry

Committee Director

James H. Yuan

Committee Member

Michael C. Beatrice

Committee Member

James R. Swanson

Call Number for Print

Special Collections LD4331.C45L55

Abstract

Protein profiles of human spermatozoa have been mapped by using high resolution two-dimensional electrophoresis which combines isoelectric focusing as the first dimension and polyacrylamide gradient gels electrophoresis in SDS as the second dimension. The profiles of normal spermatozoa! extraction, acrosomal extraction and seminal plasma showed high resolution in its separation capability. No qualitative or quantitative differences were observed by using Coomassie Brilliant Blue R-250 staining in a much higher concentration of proteins. However, coomassie blue lacks the sensitivity to detect many minor components. Therefore, a silver staining procedure was used, which resulted in greatly increased sensitivity.

Samples from normal fertile males revealed a pattern of over 200 proteins ranging in molecular weight from 15,000 to 100,000 daltons. Comparison of the mapped proteins were either by use of standard molecular markers to calculate the relative molecular weight of certain spots or by co-electrophoresis of certain known pure proteins to associate the activity with given spots.

Spermatozoa contain many different proteins mostly in trace amounts. The understanding of the distribution of these proteins may be diagnostically useful in evalution of fertility. The protein profiles of human spermatozoa were correlated with the transferrin at molecular weight 88,000 daltons, albumin at 65,000 daltons, alpha- 1-antitrypsin at 61,000 daltons, acid phosphatase at 47,000 daltons, and hyaluronidase at 26,000 daltons. There were two additional families found in spermatozoal patterns only, and the molecular weights were approximately 45,000 and 33,000 daltons.

Since proteins in the regions of 45,000, 33,000 and 26,000 daltons were missing in seminal plasma samples, proteins in theses regions are specific only for spermatozoa. No qualitative and quantitative differences were observed between spermatozoa extraction and acrosomal extraction profiles except in the region of 45,000 daltons which was missing in the acrosomal profile.

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DOI

10.25777/wjdk-jt28

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