scholarly journals Purification and characterization of testicular transferrin secreted by rat Sertoli cells

1984 ◽  
Vol 218 (2) ◽  
pp. 313-320 ◽  
Author(s):  
M K Skinner ◽  
W L Cosand ◽  
M D Griswold
Keyword(s):  
Cell Culture ◽  
Sertoli Cells ◽  
Culture Medium ◽  
Serum Transferrin ◽  
Cell Culture Medium ◽  
Purification And Characterization ◽  
Reverse Phase ◽  
Rat Serum ◽  
Molecular Masses

Sertoli cells synthesize and secrete a transferrin-like protein (testicular transferrin) [Skinner & Griswold (1980) J. Biol. Chem. 255, 1923-1925]. The purpose of the present study was to purify and characterize testicular transferrin and to compare it with serum transferrin. Testicular transferrin was obtained from the medium of cultured rat Sertoli cells, whereas serum transferrin was obtained from rat serum. Both proteins were purified with the use of phenyl-Sepharose hydrophobic chromatography and transferrin immunoaffinity chromatography. The purified proteins were shown to have similar molecular masses (75 000 Da) and amino acid compositions. The pattern of tryptic peptides from testicular and serum transferrin were found to be essentially the same when analysed by reverse-phase high-pressure liquid chromatography. The carbohydrate composition of both transferrins was determined by several colorimetric assays and g.l.c. Testicular transferrin, isolated from cell culture medium, had increased amounts of glucose, galactose and glucosamine. Serum transferrin that was incubated with cell culture medium also had a large amount of associated glucose. The results show that testicular transferrin and serum transferrin are structurally very similar and are possibly products of the same gene expressed in two different tissues, the testis and liver. However, the amount of carbohydrate associated with these two proteins is different.

Surface characterization of bone graft substitute materials conditioned in cell culture medium

2010 ◽  
Vol 42 (6-7) ◽  
pp. 452-456 ◽  
Author(s):  
Z. Mladenovic ◽  
A. Sahlin-Platt ◽  
Å. Bengtsson ◽  
M. Ransjö ◽  
A. Shchukarev
Keyword(s):  
Cell Culture ◽  
Bone Graft ◽  
Surface Characterization ◽  
Culture Medium ◽  
Bone Graft Substitute ◽  
Cell Culture Medium ◽  
Substitute Materials

scholarly journals Surface Characterization of Amorphous Zr-Al-(Ni, Cu) Alloys Immersed in Cell-Culture Medium

2002 ◽  
Vol 43 (2) ◽  
pp. 261-266 ◽  
Author(s):  
Sachiko Hiromoto ◽  
Katsuhiko Asami ◽  
An Pang Tsai ◽  
Takao Hanawa
Keyword(s):  
Cell Culture ◽  
Surface Characterization ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Cu Alloys

Characterization of fullerene colloidal suspension in a cell culture medium for in vitro toxicity assessment

2010 ◽  
Vol 6 (7) ◽  
pp. 1238 ◽  
Author(s):  
Haruhisa Kato ◽  
Naohide Shinohara ◽  
Ayako Nakamura ◽  
Masanori Horie ◽  
Katsuhide Fujita ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Colloidal Suspension ◽  
Toxicity Assessment ◽  
In Vitro Toxicity ◽  
Cell Culture Medium ◽  
A Cell ◽  
In Vitro Toxicity Assessment

Surface characterization of insulin-coated Ti6Al4V medical implants conditioned in cell culture medium: An XPS study

2017 ◽  
Vol 216 ◽  
pp. 33-38
Author(s):  
Andrey Shchukarev ◽  
Behnosh Öhrnell Malekzadeh ◽  
Maria Ransjö ◽  
Pentti Tengvall ◽  
Anna Westerlund
Keyword(s):  
Cell Culture ◽  
Surface Characterization ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Medical Implants ◽  
Xps Study

scholarly journals Human liver N-acetylglucosamine-6-sulphate sulphatase. Purification and characterization

1987 ◽  
Vol 246 (2) ◽  
pp. 347-354 ◽  
Author(s):  
C Freeman ◽  
P R Clements ◽  
J J Hopwood
Keyword(s):  
Enzyme Activity ◽  
Culture Medium ◽  
Large Subunit ◽  
Polyacrylamide Gel Electrophoresis ◽  
Purification And Characterization ◽  
Isolation And Characterization ◽  
Molecular Masses ◽  
Single Polypeptide ◽  
Column Procedure

Human N-acetylglucosamine-6-sulphate sulphatase was purified at least 50,000-fold to homogeneity in 78% yield from liver with a simple three-step four-column procedure, which consists of a concanavalin A-Sepharose/Blue A-agarose coupled step, chromatofocusing and Cu2+-chelating Sepharose chromatography. In all, four forms were isolated and partially characterized. Forms A and B, both with a pI greater than 9.5 and representing 30% and 60% respectively of the recovered enzyme activity, were separated by hydroxyapatite chromatography of the enzyme preparation obtained from the Cu2+-chelating Sepharose step. Both forms A and B had native molecular masses of 75 kDa. When analysed by SDS/polyacrylamide-gel electrophoresis, form A consists of a single polypeptide of molecular mass 78 kDa, whereas form B contained 48 kDa and 32 kDa polypeptide subunits. Neither form A nor form B was taken up from the culture medium into cultured human skin fibroblasts. The two other forms (C and D), with pI values of 5.8 and 5.4 respectively, represented approx. 7% and 3% of the total recovered enzyme activity. The native molecular masses of forms C and D were 94 kDa and approx. 75 kDa respectively. Form C contained three polypeptides with molecular masses of 48, 45 and 32 kDa. N-Acetylglucosamine-6-sulphate sulphatase activity was measured with a radiolabelled disaccharide substrate derived from heparin. The development of this substrate enabled the isolation and characterization of N-acetylglucosamine-6-sulphate sulphatase to proceed efficiently. Forms A, B and C had pH optima of 5.0, Km values of 11.7, 14.2 and 11.1 microM respectively and Vmax. values of 105, 60 and 53 nmol/min per mg of protein respectively. The molecular basis of the multiple forms of this sulphatase is not known. It is postulated that the differences in structure and properties of the four enzyme forms are due to differences in the state of processing of a large subunit.

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