Preferences help
enabled [disable] Abstract
Number of results
2009 | 56 | 3 | 455-463
Article title

Governing the monomer-dimer ratio of human cystatin c by single amino acid substitution in the hinge region

Title variants
Languages of publication
Three dimensional domain swapping is one of the mechanisms involved in formation of insoluble aggregates of some amyloidogenic proteins. It has been proposed that proteins able to swap domains may share some common structural elements like conformationally constrained flexible turns/loops. We studied the role of loop L1 in the dimerization of human cystatin C using mutational analysis. Introduction of turn-favoring residues such as Asp or Asn into the loop sequence (in position 57) leads to a significant reduction of the dimer fraction in comparison with the wild type protein. On the other hand, introduction of a proline residue in position 57 leads to efficient dimer formation. Our results confirm the important role of the loop L1 in the dimerization process of human cystatin C and show that this process can be to some extent governed by single amino acid substitution.
Physical description
  • Abrahamson M, Grubb A (1994) Increased body temperature accelerates aggregation of the Leu-68→Gln mutant cystatin C, the amyloid-forming protein in hereditary cystatin C amyloid angiopathy. Proc Natl Acad Sci 91: 1416-1420.
  • Abrahamson M, Dalboge H, Olafsson I, Carlsen S, Grubb A (1988) Efficient production of native, biologically active human cystatin C by Escherichia coli. FEBS Lett 236: 14-18.
  • Anderson WF, Ohlendorf DH, Takeda Y, Matthews BW (1981) Structure of the cro repressor from bacteriophage lambda and its interaction with DNA. Nature 290: 754-758.
  • Bennett MJ, Choe S, Eisenberg D (1994) Domain swapping: Entangling alliances between proteins. Proc Natl Acad Sci. 91: 3127-3131.
  • Bennett MJ, Sawaya MR, Eisenberg D (2006) Deposition diseases and 3D domain swapping, Structure 14: 811-824.
  • Bergdoll M, Remy M-H, Cagnon C, Masson JM, Dumas P (1996) Proline-dependent oligomerization with arm exchange. Structure 5: 391-401.
  • Bode W, Engh R, Musil D, Thiele U, Huber R, Karshikov A, Brzin J, Kos J, Turk V (1988) The 2.0 Å X-ray crystal structure of chicken egg white cystatin and its possible mode of interaction with cysteine proteinases. EMBO J 7: 2593-2599.
  • Crestfield AM, Stein WH, Moore S (1962) On the aggregation of bovine pancreatic ribonuclease. Arch Biochem Biophys 1: 217-222.
  • Dehouck Y, Biot Ch, Gilis D, Kwasigroch JM, Rooman M (2003) Sequence-structure signals of 3D domain swapping in proteins. J Mol Biol 330: 1215-1225.
  • Ding F, Prutzman KC, Campbell SL, Dokholyan NV (2006) Topological determinants of protein swapping. Structure 14: 5-14.
  • Ekiel I, Abrahamson M (1996) Folding-related dimerization of human cystatin C. J Biol Chem 271: 1314-1321.
  • Gronenborn AM (2009) Protein acrobatics in pairs - dimerization via domain swapping. Curr Opin Struct Biol 19: 39-49.
  • Henskens YM, Veerman ECI, Nieuw Amerongen AV (1996) Cystatins in health and disease. Biol Chem 377: 71-86.
  • Janowski R, Kozak M, Jankowska E, Grzonka Z, Grubb A, Abrahamson M, Jaskólski M (2001) Human cystatin C, an amyloidogenic protein dimerizes through three-dimensional domain swapping. Nat Struct Biol 8: 316-320.
  • Janowski R, Kozak M, Abrahamson M, Grubb A, Jaskólski M (2005) 3D domain-swapped human cystatin C with amyloidlike intermolecular β-sheets. Proteins 61: 570-578.
  • Jaskólski M (2001) 3D Domain swapping, protein oligomerization, and amyloid formation Acta Biochim Polon 48: 807-827.
  • Jenko Kokalj SJ, Gunčar GŠI, Morgan G, Rabzelj S, Kenig M, Staniforth RA, Waltho JP, Žerovnik E, Turk D (2007) Essential role of proline izomerization in stefin B tetramer formation. J Mol Biol 366: 1569-1579.
  • Jerala R, Žerovnik E (1999) Accessing the global minimum conformation of stefin A dimer by annealing under partially denaturating conditions. J Mol Biol 291: 1079-1098.
  • Koradi R, Billeter M, Wüthrich K (1996) MOLMOL: a program for display and analysis of macromolecular structures. J Mol Graph 14: 51-55.
  • Liu Y, Eisenberg D (2002) 3D domain swapping: As domains continue to swap. Protein Sci 11: 1285-1299.
  • London J, Skrzynia C, Goldberg ME (1974) Renaturation of Escherichia coli tryptophanase after exposure to 8 M urea. Evidence for the existence of nucleation centers. Eur J Biochem 47: 409-415.
  • Martin JR, Craven CJ, Jerala R, Kroon-Zitko L, Zerovnik E, Turk V, Waltho JP (1995) The three-dimensional solution structure of human stefin A. J Mol Biol 246: 331-343.
  • Neu HC, Heppel LA (1965) The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 240: 3685-3692.
  • Nilsson M, Wang X, Rodziewicz-Motowidło S, Janowski R, Lindstrom V, Onnerfjord P, Westermark G, Grzonka Z, Jaskolski M, Grubb A (2004) Prevention of domain swapping inhibits dimerization and amyloid fibril formation of cystatin C: use of engineered disulfide bridges, antibodies, and carboxymethylpapain to stabilize the monomeric form of cystatin C. J Biol Chem 279: 24236-24245.
  • Ptitsyn OB (1995) Molten globule and protein folding. Adv Protein Chem 47: 83-229.
  • Rodziewicz-Motowidło S, Wahlbom M, Wang X, Łągiewka J, Janowski R, Jaskólski M, Grubb A, Grzonka Z (2006) Checking the conformational stability of cystatin C and its L68Q variant by molecular dynamics studies: Why is the L68Q variant amyloidogenic? J Struct Biol 154: 68-78.
  • Rodziewicz-Motowidło S, Iwaszkiewicz J, Sosnowska R, Czaplewska P, Sobolewski E, Szymańska A, Stachowiak K, Liwo A (2009) The role of the Val57 amino-acid residue in the hinge loop of the human cystatin C. Conformational studies of the beta 2-L 1-beta3 segments of wild-type human cystatin C and its mutants. Biopolymers 91: 373-383.
  • Rousseau F, Schymkowitz JWH, Wilkinson HR, Itzhaki LS (2001) Three-dimensional domain swapping in p13suc1 occurs in the unfolded state and is controlled by conserved proline residues. Proc Natl Acad Sci USA 98: 5596-5601.
  • Rousseau F, Joost WH, Schymkowitz JWH, Itzhaki LS (2003) The unfolding story of three-dimensional domain swapping. Structure 11: 243-251.
  • Sanders A, Craven J, Higgins LD, Giannini S, Conroy MJ, Hounslow AM, Waltho JP, Staniforth RA (2004) Cystatin forms a tetramer through structural rearrangement of domain-swapped dimers prior to amyloidogenesis. J Mol Biol 336: 165-178.
  • Stachowiak K, Rodziewicz-Motowidło S, Sosnowska R, Kasprzykowski F, Łankiewicz L, Grubb A, Grzonka Z (2004) Effect of antisense peptide binding on the dimerization of human cystatin C - gel electrophoresis and molecular modeling studies. Acta Biochim Polon 51: 153-160.
  • Staniforth RA, Giannini S, Higgins LD, Conroy MJ, Hounslow AM, Jerala R, Craven CJ, Waltho JP (2001) Three-dimensional domain swapping in the folded and molten-globule states of cystatins, an amyloid-forming structural superfamily. EMBO J 20: 4774-4781.
  • Turk V, Bode W (1991) The cystatins: protein inhibitors of cysteine proteases. FEBS Lett 285: 213-219.
  • Wahlbom M, Wang X, Lindström V, Carlemalm E, Jaskolski M, Grubb A (2007) Fibrillogenic oligomers of human cystatin C are formed by propagated domain swapping. J Biol Chem 282: 18318-18326.
  • Wedemeyer WJ, Welker E, Scheraga HA (2002) Proline cis-trans isomerization and protein folding. Biochemistry 41: 14637-14644.
  • Wilmot CM, Thornton JM (1988) Analysis and prediction of the different types of β-turn in proteins. J Mol Biol 203: 221-232.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.