Ad. Due et al., A CYSTEINELESS GLUT1 GLUCOSE-TRANSPORTER HAS NORMAL FUNCTION WHEN EXPRESSED IN XENOPUS OOCYTES, Biochemical and biophysical research communications, 208(2), 1995, pp. 590-596
To test the role of cysteines in the function of GLUT1 glucose transpo
rter, site-directed mutagenesis was used to replace all six GLUT1 cyst
eines with serine residues. When the individual and combined Cys-->Ser
mutants were expressed in Xenopus laevis oocytes, zero-trans uptake o
f 3-O-methylglucose was comparable to that seen in native GLUT1. The '
'cysteineless'' construct also retained the kinetic features of GLUT1,
including an asymmetric transport mechanism and similar substrate and
inhibitor affinities. Whereas GLUT1 transport was inhibited by sulfhy
dryl reagents, that of the ''cysteineless'' construct was not. These r
esults show that cysteines are not required for GLUT1 function or olig
omer formation. The ''cysteineless'' construct may therefore serve as
a template for reintroducing cysteines back into GLUT1 at sites useful
for testing transporter structure and function. (C) 1995 Academic Pre
ss, Inc.