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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 10 (1986), S. 125-132 
    ISSN: 1573-0603
    Keywords: basic fibroblast growth factor ; heparin affinity ; hepatoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Intact human hepatoma derived basic fibroblast growth factor (bFGF) is a cationic 18 400-molecular-weight polypeptide, which stimulates the proliferation of 3T3 and endothelial cells at 1 ng/ml. bFGF has a strong affinity for heparin, and can be purified to homogeneity from human hepatoma cells using heparin-Sepharose chromatography. A three-step procedure is used: (a) extraction of the cells with 1M NaCl at pH 7.5; (b) Bio-Rex 70 cation exchange chromatography; and (c) heparin-Sepharose chromatography. The molecular weight of bFGF depends on the pH of the cell extraction step. When extracted at neutral pH, intact bFGF is obtained with a molecular weight of 18 400, however when extracted at pH 3.5 to 4.5 the molecular weight of bFGF is about 16 500. Lowering the molecular weight by acid pH extraction can be inhibited with a mixture of the proteinase inhibitors, PMSF, leupeptin, and pepstatin. These results suggest that degradation of bFGF is the result of cleavages by acid proteinases. It has been determined by amino acid sequence data and western blot analysis that the cleavages occur at the amino terminus of bFGF. The lower molecular weight form of bFGF has the same biological activity and exhibits the same chromatographic behavior on heparin-Sepharose as does intact bFGF.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 24 (1989), S. 229-236 
    ISSN: 0148-7280
    Keywords: epididymis ; sperm maturation ; sperm density ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Epididymal transit is involved in the acquisition of fertilizing ability by mammalian spermato-zoa. The epididymis is implicated in the addition and/or modification of sperm surface proteins functionally involved in the processes leading to fertilization. In order to characterize these sperm components, we have studied the modifications of sperm membrane proteins during epididymal maturation. Hamster spermatozoa were collected from the caput, corpus, and proximal and distal cauda of the epididymis and submitted to a continuous Percoll gradient centrifugation. Independently of their epididymal origin, the spermatozoa were distributed into two bands of buoyant densities of 1,045 and 1,088 on the gradient. However, the proportion of more dense spermatozoa increased progressively along the epididymis. This proportion is 87-fold higher in the distal cauda compared to the caput. SDS-PAGE electrophoresis demonstrated that although they originate from different regions of the epididymis, spermatozoa of the same density exhibited similar membrane protein patterns. In contrast, the electrophoretic patterns of spermatozoa with densities of 1,045 and 1,088 were very different One of these differences involves a 26 kD protein that is implicated in zona pellucida recognition (Sullivan and Bleau, Gamete Res 12.101-116,1985). This protein is present in dense sperm obtained from all regions of the epididymis but is absent in spermatozoa of low density. If we consider that this 26 kD protein is a ‘marker’ of surface changes occurring during sperm maturation, we can hypothesize that in the proximal segments of the epididymis a certain proportion of spermatozoa are indeed mature but that this proportion increases considerably along the epididymis.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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