Diabetes 50:1253-1262, 2001
© 2001 by the American Diabetes Association, Inc.
Hexokinase Isozyme Distribution in Human Skeletal Muscle
Vladimir B. Ritov, and
David E. Kelley
Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
Two isoforms of hexokinase (type I and type II) are expressed in skeletal muscle; however, the intracellular distribution of these hexokinase isoforms in human skeletal muscle is unclear. The current study was undertaken to assess this issue because binding of hexokinase to subcellular structures is considered to be an important mechanism in the regulation of glucose phosphorylation. Vastus lateralis muscle was obtained from healthy lean individuals. Muscle homogenate was separated at 45,000g into particulate and cytosolic fractions. The activity and subcellular distribution of hexokinase isozymes in human skeletal muscle was determined using ion-exchange chromatography and a highly sensitive high-performance liquid chromatographybased hexokinase assay. This criterion method was used to validate a modified thermal inactivation method for distinguishing type I and type II isoforms. Mean hexokinase activity was 3.88 ± 0.65 U/g wet wt or 0.64 ± 0.11 U/mU creatine kinase (CrK) in the particulate fraction and 0.45 ± 0.22 U/g wet wt or 0.07 ± 0.03 U/mU CrK in the cytosolic fraction. Hexokinase I and II accounted for 7075 and 2530% of total hexokinase activity, respectively. Nearly all (95%) of hexokinase I activity (0.52 ± 0.09 U/mU CrK) was found in the particulate fraction, consistent with the known high affinity of hexokinase I for mitochondria. Hexokinase II activity was also largely bound to the particulate fraction (72%), but 28% was found within the cytosolic fraction. Thus, within the particulate fraction, the relative contributions of hexokinase I and hexokinase II were 81 and 19%, whereas within the cytosolic fraction, the relative contributions for hexokinase I and hexokinase II were 37 and 63%.

CiteULike Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
F. G.S. Toledo, E. V. Menshikova, K. Azuma, Z. Radikova, C. A. Kelley, V. B. Ritov, and D. E. Kelley
Mitochondrial Capacity in Skeletal Muscle Is Not Stimulated by Weight Loss Despite Increases in Insulin Action and Decreases in Intramyocellular Lipid Content
Diabetes,
April 1, 2008;
57(4):
987 - 994.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
F. G.S. Toledo, E. V. Menshikova, V. B. Ritov, K. Azuma, Z. Radikova, J. DeLany, and D. E. Kelley
Effects of Physical Activity and Weight Loss on Skeletal Muscle Mitochondria and Relationship With Glucose Control in Type 2 Diabetes
Diabetes,
August 1, 2007;
56(8):
2142 - 2147.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
E. V. Menshikova, V. B. Ritov, R. E. Ferrell, K. Azuma, B. H. Goodpaster, and D. E. Kelley
Characteristics of skeletal muscle mitochondrial biogenesis induced by moderate-intensity exercise and weight loss in obesity
J Appl Physiol,
July 1, 2007;
103(1):
21 - 27.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
T. Ploug and J. Vinten
COUNTERPOINT: GLUCOSE PHOSPHORYLATION IS NOT A SIGNIFICANT BARRIER TO GLUCOSE UPTAKE BY THE WORKING MUSCLE
J Appl Physiol,
December 1, 2006;
101(6):
1805 - 1806.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
REBUTTAL FROM DRS. PLOUG AND VINTEN
J Appl Physiol,
December 1, 2006;
101(6):
1807 - 1808.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
T. Ploug and J. Vinten
Last Word: Point:Counterpoint authors respond to the commentaries on "Glucose phosphorylation is/is not a significant barrier to glucose uptake by the working muscle" that appear in this issue
J Appl Physiol,
December 1, 2006;
101(6):
1811 - 1811.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Bertoldo, R. R. Pencek, K. Azuma, J. C. Price, C. Kelley, C. Cobelli, and D. E. Kelley
Interactions Between Delivery, Transport, and Phosphorylation of Glucose in Governing Uptake Into Human Skeletal Muscle
Diabetes,
November 1, 2006;
55(11):
3028 - 3037.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
E. V. Menshikova, V. B. Ritov, L. Fairfull, R. E. Ferrell, D. E. Kelley, and B. H. Goodpaster
Effects of exercise on mitochondrial content and function in aging human skeletal muscle.
J. Gerontol. A Biol. Sci. Med. Sci.,
June 1, 2006;
61(6):
534 - 540.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
E. V. Menshikova, V. B. Ritov, F. G. S. Toledo, R. E. Ferrell, B. H. Goodpaster, and D. E. Kelley
Effects of weight loss and physical activity on skeletal muscle mitochondrial function in obesity
Am J Physiol Endocrinol Metab,
April 1, 2005;
288(4):
E818 - E825.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
V. B. Ritov, E. V. Menshikova, J. He, R. E. Ferrell, B. H. Goodpaster, and D. E. Kelley
Deficiency of Subsarcolemmal Mitochondria in Obesity and Type 2 Diabetes
Diabetes,
January 1, 2005;
54(1):
8 - 14.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. Bouche, S. Serdy, C. R. Kahn, and A. B. Goldfine
The Cellular Fate of Glucose and Its Relevance in Type 2 Diabetes
Endocr. Rev.,
October 1, 2004;
25(5):
807 - 830.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
H. A. Petersen, P. T. Fueger, D. P. Bracy, D. H. Wasserman, and A. E. Halseth
Fiber type-specific determinants of Vmax for insulin-stimulated muscle glucose uptake in vivo
Am J Physiol Endocrinol Metab,
March 1, 2003;
284(3):
E541 - E548.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. E. Kelley, J. He, E. V. Menshikova, and V. B. Ritov
Dysfunction of Mitochondria in Human Skeletal Muscle in Type 2 Diabetes
Diabetes,
October 1, 2002;
51(10):
2944 - 2950.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2001 by the American Diabetes Association.
|
|
| |
|