title: Three novel beta-propeller mutations causing Glanzmann thrombasthenia result in production of normally stable pro-alphaIIb, but variably impaired progression of pro-alphaIIbbeta3 from endoplasmic reticulum to Golgi.
abstract: BACKGROUND
  Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin alpha(IIb)beta3 (glycoprotein IIb/IIIa).
  
  
  OBJECTIVE
  To determine the molecular basis of GT and characterize the mutations by in vitro expression studies.
  
  
  METHODS
  We studied three unrelated patients from southern India whose diagnosis was consistent with GT.
  
  
  RESULTS
  Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature alpha(IIb) in the G128S mutant, in contrast to 6% and 33% of the normal amount of mature alpha(IIb) in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro-alpha(IIb) in the mutants comparable with the normal pro-alpha(IIb), but no conversion to mature alpha(IIb) in the G128S mutant, and only trace conversion to mature alpha(IIb) in the S287L and G357S mutants. The disappearance of pro-alpha(IIb) in the three mutants was similar to that in cells expressing normal alpha(IIb)beta3 or alpha(IIb) only. All three mutants demonstrated pro-alpha(IIb)beta3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker.
  
  
  CONCLUSIONS
  These three beta-propeller mutations do not affect the production of pro-alpha(IIb), its ability to complex with beta3, or its stability, but do cause variable defects in transport of pro-alpha(IIb)beta3 complexes from the endoplasmic reticulum to the Golgi.
authors.affiliation: Laboratory of Blood and Vascular Biology, The Rockefeller University, New York, NY 10021-6399, USA.
authors.forename: E J R
authors.initials: EJ
authors.lastname: Nelson
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: W B
authors.initials: WB
authors.lastname: Mitchell
authors.forename: M
authors.initials: M
authors.lastname: Chandy
authors.forename: A
authors.initials: A
authors.lastname: Srivastava
authors.forename: B S
authors.initials: BS
authors.lastname: Coller
mesh_terms: D000328:Adult; D002675:Child, Preschool; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005260:Female; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D008297:Male; D015394:Molecular Structure; D020125:Mutation, Missense; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D040201:Platelet Membrane Glycoprotein IIb / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011498:Protein Precursors; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000235:genetics; D013915:Thrombasthenia / Q000235:genetics*
chemical_list: D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D040201:Platelet Membrane Glycoprotein IIb; D011498:Protein Precursors