Synthesis in E. coli of alpha 1-antitrypsin variants of therapeutic potential for emphysema and thrombosis. Academic Article uri icon

Overview

MeSH

  • Amino Acid Sequence
  • Emphysema
  • Escherichia coli
  • Genetic Engineering
  • Humans
  • Mutation
  • Pancreatic Elastase
  • Structure-Activity Relationship
  • Thrombin
  • Thrombosis

MeSH Major

  • alpha 1-Antitrypsin

abstract

  • The primary function of alpha 1-antitrypsin (alpha 1-AT), an antiprotease produced by the liver, is the inhibition of neutrophil elastase, a protease capable of hydrolysing most connective tissue components. The importance of alpha 1-AT is demonstrated by the high incidence of early-onset emphysema in individuals with hereditary alpha 1-AT deficiency (Type PiZZ), in whom serum levels of alpha 1-AT are 10-20% of normal. Oxidants in tobacco smoke can inactivate alpha 1-AT in vitro, and studies have shown that alpha 1-AT from the lungs of individuals who smoke cigarettes may also be partially inactivated, perhaps explaining the high incidence of emphysema associated with cigarette smoking. Oxidative inactivation is probably due to modification of the Met residue (Met358) at the P1 subsite position of the elastase binding site of the protein. To study the possibility of modulating the biological properties of alpha 1-AT, we have introduced selected sequence modifications at the reactive site by in vitro mutation of a cloned alpha 1-AT complementary DNA. We describe here the characterization of two alpha 1-AT analogues produced in Escherichia coli. The first, alpha 1-AT(Met385----Val), is not only fully active as an elastase inhibitor but is also resistant to oxidative inactivation. The other, alpha 1-AT(Met358----Arg), no longer inhibits elastase but is an efficient thrombin inhibitor. The active site of the latter is identical to that of the alpha 1-AT (Pittsburgh) variant, which was associated with a fatal bleeding disorder.

publication date

  • January 10, 1985

has subject area

  • Amino Acid Sequence
  • Emphysema
  • Escherichia coli
  • Genetic Engineering
  • Humans
  • Mutation
  • Pancreatic Elastase
  • Structure-Activity Relationship
  • Thrombin
  • Thrombosis
  • alpha 1-Antitrypsin

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed ID

  • 3880873

Additional Document Info

start page

  • 149

end page

  • 151

volume

  • 313

number

  • 5998