Spontaneous asparaginyl deamidation of canine milk lysozyme under mild conditions

Proteins
Yasuhiro NonakaKeiichi Kawano

Abstract

Asparaginyl deamidation is a common form of nonenzymatic degradation of proteins and peptides. As it introduces a negative charge spontaneously and irreversibly, charge heterogeneity can be accumulated in protein solution during purification, preservation, and experiments. In this study, canine milk lysozyme (CML), a useful model for the study of the molten globule state, exhibited charge heterogeneity after sample purification. Four Asn residues in CML deamidated rapidly under mild conditions: pH 8.0 and 30 degrees C. Other than these residues, one Asn residue, which was stable in the native state, was labile to deamidation in the unfolded state. This suggests that the structural formation around Asn can suppress deamidation. Substitutions of these labile Asn residues to Gln residues prevented deamidation effectively. Because the substitutions did not disrupt the structural formation of the native and molten globule states, they will enable more precise analyses for physical and structural studies.

References

Jan 1, 1990·Annual Review of Biochemistry·P S Kim, R L Baldwin
Jul 1, 1986·International Journal of Peptide and Protein Research·Y C MeinwaldH A Scheraga
Sep 1, 1986·International Journal of Peptide and Protein Research·C SecchiS Ronchi
Jun 14, 1985·Science·T J Ahern, A M Klibanov
Jan 1, 1974·Current Topics in Cellular Regulation·A B Robinson, C J Rudd
Jul 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·C F Midelfort, A H Mehler
Oct 1, 1981·Journal of Biochemistry·T ImotoD Koga
Jan 1, 1996·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·K Kuwajima
Nov 24, 1999·The Journal of Peptide Research : Official Journal of the American Peptide Society·S Capasso, S Salvadori
Dec 14, 1999·Protein Science : a Publication of the Protein Society·A A KoskyD N Brems
Mar 22, 2000·The Journal of Peptide Research : Official Journal of the American Peptide Society·S Capasso
Sep 28, 2000·The Journal of Peptide Research : Official Journal of the American Peptide Society·M XieR L Schowen
Nov 28, 2000·Acta Crystallographica. Section D, Biological Crystallography·A Vagin, A Teplyakov
Feb 7, 2001·Proceedings of the National Academy of Sciences of the United States of America·N E Robinson, A B Robinson
Apr 11, 2001·Proceedings of the National Academy of Sciences of the United States of America·N E Robinson, A B Robinson
Jan 16, 2002·Protein Science : a Publication of the Protein Society·Melanie R NilssonDaniel P Raleigh
Apr 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Noah E Robinson
Feb 20, 2003·Protein Science : a Publication of the Protein Society·Herman Van DaelMarcel Joniau
Mar 5, 2003·Pharmacological Reviews·Michael R Yeaman, Nannette Y Yount
May 14, 2004·The Journal of Peptide Research : Official Journal of the American Peptide Society·N E RobinsonA B Robinson
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

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Citations

Apr 27, 2011·The Journal of Biological Chemistry·Quanzhou LuoJette Wypych
Jul 18, 2009·Biologicals : Journal of the International Association of Biological Standardization·Y Diana LiuGregory C Flynn
Jun 24, 2008·Protein Science : a Publication of the Protein Society·Takumi TakataKirsten J Lampi
Apr 23, 2019·Food Chemistry·Saliha Si Ahmed ZenniaJean-Michel Girardet

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