Cardiac alpha-crystallin |
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Authors: | Stefano Longoni Sari Lattonen Gillian Bullock Michele Chiesi |
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Institution: | (1) Department Pharmaceutical Research, Ciba-Geigy, CH-4002 Basel, Switzerland;(2) Present address: Institut Dr. Viollier, Spalenring 145, CH-4002 Basel, Switzerland |
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Abstract: | Summary A major component of the soluble fraction of rat heart is a homopolymer (Mr about 400–650 k) of a small protein (Mr about 20 k). This cardiac protein, which is highly homologous to alpha-B-crystallin, was isolated in its native state and visualized by electron microscopy. A homogeneous population of globular particles with an average diameter of about 14-16 nM could be seen using either negative staining or rotary shadowing procedures. The structures were globular in nature with a central depression (torus-like structures). Polyclonal antibodies, raised against the cardiac crystallin, were used for the immunocytochemical localization of the macromolecular complexes. Using fluorescent secondary antibodies, a clear and sharp striation of fixed and permeabilized rat heart myocytes could be observed, similar to that observed with anti-desmin antibodies and characteristic for the central region of the I-band. Cardiac crystallin was not found associated with F-actin in preparations of rat heart myofibrils. On the other hand, it was a major contaminant of cardiac desmin preparations. These observations suggest that cardiac crystallin is involved in the organization of cytoskeletal filaments of the Z-lines.Abbreviations SDS
Sodium Dodecyl Sulfate
- SDS-PAGE
SDS-Polyacrylamide Gel Electrophoresis
- PMSF
Phenyl Methyl Sulfonyl Fluoride
- MOPS
3-Morpholino Propanesulfonic Acid
- TX-100
Octyl Phenoxy Polyethoxyethanol
- CHAPS
(3-((3-Cholamidopropyl)Dimethylammonio)-1-Propanesulfonate),
- BSA
Bovine Serum Albumin
- FITC
Fluorescein Isothiocyanate
- PO
Peroxidase |
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Keywords: | heart alpha-crystallin Z-line quaternary structure immunocytochemical localization microscopy |
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