Monoclonal antibodies to rabbit hepatocyte myosin that cross-react with human liver myosin.
We prepared polyclonal and monoclonal antibodies against myosin purified from rabbit hepatocytes. Immunoblotting analyses revealed that the polyclonal antibody and four (HM1, HM2, HM3 and HM4) of five monoclonal antibodies reacted with myosin heavy chain. Chymotryptic cleavage of myosin yielded a 130 kDa fragment comprising the tail portion of the myosin heavy chain and a 67 kDa fragment comprising the ATP-binding active site of the myosin head. All active antibodies reacted with epitopes localized in the 130 kDa fragment. Monoclonal antibodies HM3 and HM4 and the polyclonal antibody reacted strongly with myosin heavy chains from a human liver homogenate prepared from a surgically resected liver specimen. Immunocytochemical analyses showed that myosin is localized along the plasma membrane as well as around the bile canaliculi in both rabbit and human hepatocytes. Immunocytochemical analyses on liver blocks obtained from those patients who suffered various types of diseases accompanying cholestasis clearly indicated a marked increase in pericanalicular myosin. This altered myosin localization is analogous to that observed in phalloidin-treated liver. Thus, myosin localization, determined using these antibodies, can provide a valid morphological basis for diagnosing the pathological state of the patient liver.
Ueno
T
T
Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan.
Watanabe
S
S
Hirose
M
M
Nozawa
R
R
Yasuura
S
S
Egawa
H
H
Sato
N
N
Kominami
E
E
D000328:Adult; D000368:Aged; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007223:Infant; D008099:Liver / Q000378:metabolism*; D008107:Liver Diseases / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D018995:Myosin Heavy Chains / Q000276:immunology; D015879:Myosin Subfragments / Q000276:immunology; D009218:Myosins / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015879:Myosin Subfragments; D018995:Myosin Heavy Chains; D009218:Myosins