Purification and characterization of a major zinc binding protein from renal brush border membrane of rat.
In spite of the fact that zinc is an essential trace element, mechanisms that contribute to zinc homeostasis in mammals are poorly understood. An attempt has been made to identify and purify zinc binding components from renal brush border membrane (BBM), which could be involved in the binding of zinc and the subsequent translocation across the BBM. A 40 kDa major zinc binding protein has been identified and purified from renal BBM, which showed a dissociation constant (Kd) of 211 microM and maximal binding (Bmax) of 207 nmol/mg protein. 8 g zinc atoms could interact with 1 mol of protein. Specificity of the protein for zinc was checked by metal displacement and UV-absorption assay. It was found that only Cd2+ could displace the zinc bound to the protein. Other metals tested (Cu2+, Mg2+, Ca2+) did not show any interaction with the protein. These data indicated that purified protein is highly specific and has a high affinity for zinc. The carbohydrate content was found to be 7.85 mg% in the purified protein. Immunofluorescence localization of this protein in kidney sections revealed that this major zinc binding protein is exclusively localized in the proximal convoluted tubules. These results suggested that the 40 kDa major zinc binding transmembrane glycoprotein is highly specific for zinc and has a high affinity for zinc.
Kumar
R
R
Department of Biochemistry, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India.
Prasad
R
R
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D007684:Kidney Tubules / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008871:Microvilli / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000378:metabolism*
D002352:Carrier Proteins; D008562:Membrane Glycoproteins; C017800:zinc-binding protein; D015032:Zinc