Low and High Km Forms of R;Regular Article Dinitrophenylglutathione-stimulated ATPase in Bovine Lens

Kshama Kumari; Naseem H. Ansari; Manju Saxena; Yogesh C. Awasthi; Satish K. Srivastava

doi:10.1006/exer.1993.1120

Low and High K_m Forms of R;Regular Article Dinitrophenylglutathione-stimulated ATPase in Bovine Lens

Kshama Kumari, Naseem H. Ansari, Manju Saxena, Yogesh C. Awasthi, Satish K. Srivastava

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Dinitrophenyl S-glutathione (Dnp-SG) ATPase which catalyses the hydrolysis of ATP in the presence of GSH-conjugates has been implicated previously in the transport of these conjugates. In the present studies we demonstrate that Dnp-SG ATPase is present in bovine lens epithelium and cortex. The specific activity per mg membrane protein was found to be 75-fold higher in the epithelium as compared to the cortex. No enzyme was detected in the nuclear region of the lens. Dnp-SG ATPase was purified from bovine lens epithelium and cortex using Dnp-SG-Sepharose 6MB affinity chromatography. The partially purified Dnp-SG ATPase had two distinct K_m values, 120 μM and 1·0 mM. The antibodies raised against human erythrocyte Dnp-SG ATPase cross-reacted with the bovine lens epithelium Dnp-SG ATPase which was identified by Western blot as a band corresponding to an approximate M_r value of 80000 Da.

Original language	English (US)
Pages (from-to)	243-247
Number of pages	5
Journal	Experimental Eye Research
Volume	57
Issue number	2
DOIs	https://doi.org/10.1006/exer.1993.1120
State	Published - Aug 1993
Externally published	Yes

Keywords

Bovine lens
Dnp-SG ATPase
Glutathione
Glutathione conjugates
Xenobiotic transport

ASJC Scopus subject areas

Ophthalmology
Sensory Systems
Cellular and Molecular Neuroscience

Access to Document

10.1006/exer.1993.1120

Cite this

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title = "Low and High Km Forms of R;Regular Article Dinitrophenylglutathione-stimulated ATPase in Bovine Lens",

abstract = "Dinitrophenyl S-glutathione (Dnp-SG) ATPase which catalyses the hydrolysis of ATP in the presence of GSH-conjugates has been implicated previously in the transport of these conjugates. In the present studies we demonstrate that Dnp-SG ATPase is present in bovine lens epithelium and cortex. The specific activity per mg membrane protein was found to be 75-fold higher in the epithelium as compared to the cortex. No enzyme was detected in the nuclear region of the lens. Dnp-SG ATPase was purified from bovine lens epithelium and cortex using Dnp-SG-Sepharose 6MB affinity chromatography. The partially purified Dnp-SG ATPase had two distinct Km values, 120 μM and 1·0 mM. The antibodies raised against human erythrocyte Dnp-SG ATPase cross-reacted with the bovine lens epithelium Dnp-SG ATPase which was identified by Western blot as a band corresponding to an approximate Mr value of 80000 Da.",

keywords = "Bovine lens, Dnp-SG ATPase, Glutathione, Glutathione conjugates, Xenobiotic transport",

author = "Kshama Kumari and Ansari, {Naseem H.} and Manju Saxena and Awasthi, {Yogesh C.} and Srivastava, {Satish K.}",

year = "1993",

month = aug,

doi = "10.1006/exer.1993.1120",

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volume = "57",

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AU - Kumari, Kshama

AU - Ansari, Naseem H.

AU - Saxena, Manju

AU - Awasthi, Yogesh C.

AU - Srivastava, Satish K.

PY - 1993/8

Y1 - 1993/8

N2 - Dinitrophenyl S-glutathione (Dnp-SG) ATPase which catalyses the hydrolysis of ATP in the presence of GSH-conjugates has been implicated previously in the transport of these conjugates. In the present studies we demonstrate that Dnp-SG ATPase is present in bovine lens epithelium and cortex. The specific activity per mg membrane protein was found to be 75-fold higher in the epithelium as compared to the cortex. No enzyme was detected in the nuclear region of the lens. Dnp-SG ATPase was purified from bovine lens epithelium and cortex using Dnp-SG-Sepharose 6MB affinity chromatography. The partially purified Dnp-SG ATPase had two distinct Km values, 120 μM and 1·0 mM. The antibodies raised against human erythrocyte Dnp-SG ATPase cross-reacted with the bovine lens epithelium Dnp-SG ATPase which was identified by Western blot as a band corresponding to an approximate Mr value of 80000 Da.

AB - Dinitrophenyl S-glutathione (Dnp-SG) ATPase which catalyses the hydrolysis of ATP in the presence of GSH-conjugates has been implicated previously in the transport of these conjugates. In the present studies we demonstrate that Dnp-SG ATPase is present in bovine lens epithelium and cortex. The specific activity per mg membrane protein was found to be 75-fold higher in the epithelium as compared to the cortex. No enzyme was detected in the nuclear region of the lens. Dnp-SG ATPase was purified from bovine lens epithelium and cortex using Dnp-SG-Sepharose 6MB affinity chromatography. The partially purified Dnp-SG ATPase had two distinct Km values, 120 μM and 1·0 mM. The antibodies raised against human erythrocyte Dnp-SG ATPase cross-reacted with the bovine lens epithelium Dnp-SG ATPase which was identified by Western blot as a band corresponding to an approximate Mr value of 80000 Da.

KW - Bovine lens

KW - Dnp-SG ATPase

KW - Glutathione

KW - Glutathione conjugates

KW - Xenobiotic transport

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