title: Abnormal thiol reactivity of tropomyosin in essential hypertension and its association with abnormal sodium-lithium countertransport kinetics.
abstract: OBJECTIVE
  To identify a thiol protein that is abnormal in a subgroup of essential hypertensive (EHT) patients who have a strong family history of hypertension and cardiovascular disease and have a low Km of erythrocyte Na/Li countertransport (CT).
  
  
  METHODS
  To detect biotin maleimide labelling of a key thiol protein to investigate its reaction with N-ethylmaleimide (NEM) in normal and EHT erythrocytes.
  
  
  RESULTS
  The thiol protein of 33 kDa apparent molecular weight (p33) identified by the loss of labelling with biotin maleimide was identified as tropomyosin due to its retarded running in 6 mol/l urea gels and immunoblotting. The NEM reaction with p33 detected by loss of subsequent biotin maleimide labelling is biphasic in normal control erythrocytes with the rate in the first 30 s double that after 30 s. In EHT erythrocytes NEM reaction (1) after 30 s is faster than normal and (2) in the first 30 s causes a paradoxical increase in apparent biotin maleimide labelling. In normal control erythrocytes, the loss of biotin maleimide labelling with NEM reaction or the faster phenylmaleimide reaction follows the same time course as the decrease in Km of Na/Li CT.
  
  
  CONCLUSIONS
  NEM reaction with p33 requires two thiols. Only the cytoskeletal form of tropomyosin from the TM3 gene has more than one thiol group and agrees with SDS-PAGE mobility. Tropomyosin is a strong candidate to explain the familial abnormality in EHT with abnormal Na/ Li CT and it could explain many of the characteristics of this disease.
authors: Department of Medicine, School of Medicine, University of Newcastle-upon-Tyne, UK.
authors: S L
authors: SL
authors: Watkins
authors: I C
authors: IC
authors: West
authors: R
authors: R
authors: Wilkinson
authors: T H
authors: TH
authors: Thomas
mesh_terms: D001710:Biotin; D016022:Case-Control Studies; D004912:Erythrocytes / Q000378:metabolism; D005033:Ethylmaleimide; D005260:Female; D006801:Humans; D006973:Hypertension / Q000097:blood* / Q000235:genetics; D017136:Ion Transport; D007700:Kinetics; D008094:Lithium / Q000097:blood*; D008297:Male; D012964:Sodium / Q000097:blood*; D013438:Sulfhydryl Compounds / Q000097:blood; D013439:Sulfhydryl Reagents; D014335:Tropomyosin / Q000097:blood* / Q000737:chemistry / Q000235:genetics
chemical_list: D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D014335:Tropomyosin; D001710:Biotin; D008094:Lithium; D012964:Sodium; D005033:Ethylmaleimide