Activation of calpain in myocardial infarction: An immunohistochemical study using a calpain antibody raised against active site histidine- containing peptide

Mitoshi Kunimatsu, Toyohiro Tada, Yasuko Narita, Yasuhiko Ozaki, Zhen Qui Liu, Thomas (Tom) Shearer, Makoto Sasaki

Research output: Contribution to journalArticle

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Abstract

Tissue damage resulting from ischemia due to myocardial infarction is thought to be intensified by the proteolytic action of endogenous enzymes. Calpain (calcium dependent cysteine protease) is considered to be a highly likely candidate, since it is activated by calcium ion which increases in concentration under conditions of ischemia. We prepared a mono-specific antibody against the active site histidine stretch, Lys-Leu-Val-Lys-Gly-His- Ala-Tyr-Ser-Val, in the calpain 80 kDa large subunit. The specificity of the antibody was verified by its inhibitory effect on the caseinolytic activity of both μ- and m-calpains, western blotting analysis, and by absorption with the antigen peptide. The antibody was used to localize the intracellular distribution of activated calpains in infarcted regions of the human heart. The results showed that myocardial cells affected by ischemia were stained by the antibody, allowing damaged cells to be distinguished from cells of unaffected regions and that the immunostained regions were essentially the same regions as those identified by dense eosinophilic staining with hematoxylin and eosin. However, the staining pattern obtained with the antibody, was characteristic in denser staining at the cell periphery, whereas the damaged cells were stained homogeneously by hematoxylin and eosin. By the former method, results of staining indicated that the activation site of the calpain proenzyme was in the peri-plasma membrane, whereas by the latter method, diffusely distributed plasma proteins such as albumin and immunoglobulins were visualized as demonstrated in earlier reports.

Original languageEnglish (US)
Pages (from-to)7-15
Number of pages9
JournalCardiovascular Pathology
Volume8
Issue number1
DOIs
StatePublished - Jan 1999

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Calpain
Histidine
Catalytic Domain
Myocardial Infarction
Peptides
Antibodies
Staining and Labeling
Ischemia
Hematoxylin
Eosine Yellowish-(YS)
Calcium
Enzyme Precursors
Antibody Specificity
Cysteine Proteases
Immunoglobulins
Blood Proteins
Albumins
Western Blotting
Cell Membrane
Ions

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Pathology and Forensic Medicine

Cite this

Activation of calpain in myocardial infarction : An immunohistochemical study using a calpain antibody raised against active site histidine- containing peptide. / Kunimatsu, Mitoshi; Tada, Toyohiro; Narita, Yasuko; Ozaki, Yasuhiko; Liu, Zhen Qui; Shearer, Thomas (Tom); Sasaki, Makoto.

In: Cardiovascular Pathology, Vol. 8, No. 1, 01.1999, p. 7-15.

Research output: Contribution to journalArticle

Kunimatsu, Mitoshi ; Tada, Toyohiro ; Narita, Yasuko ; Ozaki, Yasuhiko ; Liu, Zhen Qui ; Shearer, Thomas (Tom) ; Sasaki, Makoto. / Activation of calpain in myocardial infarction : An immunohistochemical study using a calpain antibody raised against active site histidine- containing peptide. In: Cardiovascular Pathology. 1999 ; Vol. 8, No. 1. pp. 7-15.
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