Abstract |
High levels of alpha B-crystallin are present in the cardiomyocyte, yet little is understood about the function and importance of this protein. Like many other small heat shock proteins, alpha B-crystallin forms large oligomeric complexes whose size can be regulated by posttranslational modifications. The size of these complexes can modify the function of the protein. A naturally occurring COOH-terminal mutant has many detrimental effects in the lens of the eye and altered oligomerization. Therefore, we mutated the two COOH-terminal lysines of alpha B-crystallin to glycines (K174/175G) and adenovirally mounted them to transduce cardiomyocytes. We analyzed the effect of this mutation on oligomerization, microtubular stabilization, and ischemic outcome. A nearly 45% downward shift in complex size was observed with the mutant by native PAGE followed by immunoblotting. The overexpressed protein no longer protected the tubulin cytoskeleton against ischemic stress by confocal analysis. The mutant caused a 30% increase in cytosolic enzyme release with ischemia compared with control, whereas a 33% decrease was associated with wild-type alpha B-crystallin overexpression. We conclude that the COOH terminus of alpha B-crystallin is crucial to its proper function.
|
Authors | Jody L Martin, Wolfgang F Bluhm, Huaping He, Ruben Mestril, Wolfgang H Dillmann |
Journal | American journal of physiology. Heart and circulatory physiology
(Am J Physiol Heart Circ Physiol)
Vol. 283
Issue 1
Pg. H85-91
(Jul 2002)
ISSN: 0363-6135 [Print] United States |
PMID | 12063278
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
|
Chemical References |
- Crystallins
- Macromolecular Substances
- Tubulin
- Lysine
|
Topics |
- Animals
- Cells, Cultured
- Crystallins
(biosynthesis, genetics)
- Cytoskeleton
(metabolism)
- Electrophoresis, Polyacrylamide Gel
- Immunoblotting
- Lysine
(genetics)
- Macromolecular Substances
- Microtubules
(metabolism)
- Mutagenesis, Site-Directed
- Myocardial Ischemia
(metabolism)
- Myocardium
(cytology, metabolism)
- Rats
- Structure-Activity Relationship
- Transduction, Genetic
- Tubulin
(metabolism)
|