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Cardiac alpha-crystallin

II. Intracellular localization

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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.

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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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Correspondence to Michele Chiesi.

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Longoni, S., Lattonen, S., Bullock, G. et al. Cardiac alpha-crystallin. Mol Cell Biochem 97, 121–128 (1990). https://doi.org/10.1007/BF00221053

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Key words

  • heart
  • alpha-crystallin
  • Z-line
  • quaternary structure
  • immunocytochemical localization
  • microscopy