Regulation of the human Na+-dependent glucose cotransporter hSGLT2

被引:92
作者
Ghezzi, Chiara [1 ]
Wright, Ernest M. [1 ]
机构
[1] Univ Calif Los Angeles, David Geffen Sch Med, Dept Physiol, Los Angeles, CA 90095 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 2012年 / 303卷 / 03期
基金
美国国家卫生研究院; 瑞士国家科学基金会;
关键词
protein kinases; insulin; kidney; AMINOBUTYRIC-ACID TRANSPORTER; PROTEIN-KINASE; ACTIVATION; MODULATION; EXPRESSION; KINETICS;
D O I
10.1152/ajpcell.00115.2012
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Ghezzi C, Wright EM. Regulation of the human Na+-dependent glucose cotransporter hSGLT2. Am J Physiol Cell Physiol 303: C348-C354, 2012. First published June 6, 2012; doi: 10.1152/ajpcell.00115.2012.-The human Na+-glucose cotransporter SGLT2 is expressed mainly in the kidney proximal convoluted tubule where it is considered to be responsible for the bulk of glucose reabsorption. Phosphorylation profiling has revealed that SGLT2 exists in a phosphorylated state in the rat renal proximal tubule cortex, so we decided to investigate the regulation of human SGLT2 (hSGLT2) by protein kinases. hSGLT2 was expressed in human embryonic kidney (HEK) 293T cells, and the activity of the protein was measured using radiotracer and whole cell patch-clamp electrophysiology assays before and after activation of protein kinases. 8-Bromo-adenosine cAMP (8-Br-cAMP) was used to activate protein kinase A, and sn-1,2-dioctanoylglycerol (DOG) was used to activate protein kinase C (PKC). 8-Br-cAMP stimulated D-[alpha-methyl-C-14] glucopyranoside ([C-14]alpha-MDG) uptake and Na+-glucose currents by 200% and DOG increased [C-14]alpha-MDG uptake and Na+-glucose currents by 50%. In both cases the increase in SGLT2 activity was marked by an increase in the maximum rate of transport with no change in glucose affinity. These effects were completely negated by mutation of serine 624 to alanine. Insulin induced a 250% increase in Na+-glucose transport by wild-type but not S624A SGLT2. Parallel studies confirmed that the activity of hSGLT1 was regulated by PKA and PKC due to changes in the number of transporters in the cell membrane. hSGLT1 was relatively insensitive to insulin. We conclude that hSGLT1 and hSGLT2 are regulated by different mechanisms and suggest that insulin is an SGLT2 agonist in vivo.
引用
收藏
页码:C348 / C354
页数:7
相关论文
共 29 条
  • [1] BLUMENTHAL EM, 1992, J NEUROSCI, V12, P290
  • [2] Butlen D, 1988, PFLUGERS ARCH, V29, P325
  • [3] COREY JL, 1994, J BIOL CHEM, V269, P14759
  • [4] EDELMAN AM, 1987, ANNU REV BIOCHEM, V56, P567, DOI 10.1146/annurev.biochem.56.1.567
  • [5] EFFECT OF DIABETES AND INSULIN ON THE MAXIMUM CAPACITY OF THE RENAL TUBULES TO REABSORB GLUCOSE
    FARBER, SJ
    BERGER, EY
    EARLE, DP
    [J]. JOURNAL OF CLINICAL INVESTIGATION, 1951, 30 (02) : 125 - 129
  • [6] Large-scale phosphoproteomic analysis of membrane proteins in renal proximal and distal tubule
    Feric, Marina
    Zhao, Boyang
    Hoffert, Jason D.
    Pisitkun, Trairak
    Knepper, Mark A.
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2011, 300 (04): : C755 - C770
  • [7] Electrophysiological characterization of the flounder type II Na+/P-i cotransporter (NaPi-5) expressed in Xenopus laevis oocytes
    Forster, IC
    Wagner, CA
    Busch, AE
    Lang, F
    Biber, J
    Hernando, N
    Murer, H
    Werner, A
    [J]. JOURNAL OF MEMBRANE BIOLOGY, 1997, 160 (01) : 9 - 25
  • [8] Protein kinase C activators induce membrane retrieval of type IINa+-phosphate cotransporters expressed in Xenopus oocytes
    Forster, IC
    Traebert, M
    Jankowski, M
    Stange, G
    Biber, J
    Murer, H
    [J]. JOURNAL OF PHYSIOLOGY-LONDON, 1999, 517 (02): : 327 - 340
  • [9] PROTEIN-KINASE-C CONSENSUS SITES AND THE REGULATION OF RENAL NA/P-I-COTRANSPORT (NAPI-2) EXPRESSED IN XENOPUS-LAEVIS OOCYTES
    HAYES, G
    BUSCH, AE
    LANG, F
    BIBER, J
    MURER, H
    [J]. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1995, 430 (05): : 819 - 824
  • [10] HEI YJ, 1991, MOL PHARMACOL, V39, P233