Functional consequences and rescue potential of pathogenic missense mutations in tripeptidyl peptidase I

Hum Mutat. 2010 Jun;31(6):710-21. doi: 10.1002/humu.21251.


There are 35 missense mutations among 68 different mutations in the TPP1 gene, which encodes tripeptidyl peptidase I (TPPI), a lysosomal aminopeptidase associated with classic late-infantile neuronal ceroid lipofuscinosis (CLN2 disease). To elucidate the molecular mechanisms underlying TPPI deficiency in patients carrying missense mutations and to test the amenability of mutant proteins to chemical chaperones and permissive temperature treatment, we introduced individually 14 disease-associated missense mutations into human TPP1 cDNA and analyzed the cell biology of these TPPI variants expressed in Chinese hamster ovary cells. Most TPPI variants displayed obstructed transport to the lysosomes, prolonged half-life of the proenzyme, and residual or no enzymatic activity, indicating folding abnormalities. Protein misfolding was produced by mutations located in both the prosegment (p.Gly77Arg) and throughout the length of the mature enzyme. However, the routes of removal of misfolded proteins by the cells varied, ranging from their efficient degradation by the ubiquitin/proteasome system to abundant secretion. Two TPPI variants demonstrated enhanced processing in response to folding improvement treatment, and the activity of one of them, p.Arg447His, showed a fivefold increase under permissive temperature conditions, which suggests that folding improvement strategies may ameliorate the function of some misfolding TPPI mutant proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminopeptidases / genetics*
  • Aminopeptidases / metabolism*
  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / genetics*
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / metabolism*
  • Endoplasmic Reticulum / metabolism
  • Enzyme Stability
  • Genetic Complementation Test
  • Humans
  • Immunoblotting
  • Infant
  • Lysosomes / metabolism
  • Microscopy, Confocal
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism
  • Mutant Proteins / metabolism
  • Mutation, Missense*
  • Neuronal Ceroid-Lipofuscinoses / enzymology
  • Neuronal Ceroid-Lipofuscinoses / genetics
  • Protein Transport
  • Serine Proteases / genetics*
  • Serine Proteases / metabolism*
  • Temperature
  • Time Factors
  • Transfection


  • Molecular Chaperones
  • Mutant Proteins
  • Serine Proteases
  • Aminopeptidases
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
  • tripeptidyl-peptidase 1