Camel kidney ferritin: isolation and partial characterization.
Camel kidney ferritin was isolated from a tissue homogenate by thermal denaturation, ammonium sulphate fractionation, Sephacryl S-300 gel filtration and DEAE-blue gel affinity chromatography. The yield and the iron and neutral carbohydrate contents were 0.012 mg/g wet tissue, 4.0% and 2.7%, respectively. The phosphate:iron ratio was 0.13, twofold lower than that reported for camel liver ferritin. Native gel electrophoresis revealed the presence of a monomeric ferritin. SDS gel electrophoresis and immunoblotting showed two types of subunits, heavy and light, contrary to the extensive heterogeneity observed in camel liver ferritin. In general, the tissue ferritins shared a similar amino acid composition. However, a twofold lower glycine and an eightfold higher arginine content were recorded for camel kidney ferritin. In addition, kidney ferritin had a relatively high content of glutamic acid. Cross-reactivity studies by Ouchterlony double diffusion and noncompetitive indirect ELISA revealed a distinct cross-reactivity between buffalo ferritin antiserum and camel liver ferritin, but camel liver ferritin showed only weak cross-reactivity.
Sobha
G
G
Department of Biochemistry, University College of Science, Osmania University, Andhra Pradesh, India.
Suryakala
S
S
Geetha
C
C
Deshpande
V
V
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002162:Camelus / Q000502:physiology*; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity / Q000662:veterinary; D002850:Chromatography, Gel / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005293:Ferritins / Q000737:chemistry / Q000302:isolation & purification*; D015151:Immunoblotting / Q000662:veterinary; D005779:Immunodiffusion / Q000662:veterinary; D007501:Iron / Q000032:analysis; D007668:Kidney / Q000737:chemistry*; D008099:Liver / Q000737:chemistry; D011506:Proteins / Q000032:analysis; D013054:Spectrophotometry, Atomic / Q000662:veterinary
D000596:Amino Acids; D002241:Carbohydrates; D011506:Proteins; D005293:Ferritins; D007501:Iron