Abstract:
:Glucose-6-phosphatase is a multicomponent system located in the endoplasmic reticulum, involving both a catalytic subunit (G6PC) and several substrate and product carriers. The glucose-6-phosphate carrier is called G6PT1. Using light scattering, we determined K(D) values for phosphate and glucose transport in rat liver microsomes (45 and 33mM, respectively), G6PT1 K(D) being too low to be estimated by this technique. We provide evidence that phosphate transport may be carried out by an allosteric multisubunit translocase or by two distinct proteins. Using chemical modifications by sulfhydryl reagents with different solubility properties, we conclude that in G6PT1, one thiol group important for activity is facing the cytosol and could be Cys(121) or Cys(362). Moreover, a different glucose-6-phosphate translocase, representing 20% of total glucose-6-phosphate transport and insensitive to N-ethylmaleimide modification, could coexist with liver G6PT1. In the G6PC protein, an accessible thiol group is facing the cytosol and, according to structural predictions, could be Cys(284).
journal_name
Arch Biochem Biophysjournal_title
Archives of biochemistry and biophysicsauthors
Clottes E,Middleditch C,Burchell Adoi
10.1016/s0003-9861(02)00523-4subject
Has Abstractpub_date
2002-12-01 00:00:00pages
33-41issue
1eissn
0003-9861issn
1096-0384pii
S0003986102005234journal_volume
408pub_type
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