Skip Navigation


Journal of Biochemistry Advance Access originally published online on March 15, 2008
Journal of Biochemistry 2008 144(1):21-32; doi:10.1093/jb/mvn037
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplementary Data
Right arrow All Versions of this Article:
144/1/21    most recent
mvn037v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Biswas, A.
Right arrow Articles by Nagaraj, R. H.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Biswas, A.
Right arrow Articles by Nagaraj, R. H.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2008 The Japanese Biochemical Society

Chemical Modulation of the Chaperone Function of Human {alpha}A-Crystallin

Ashis Biswas1,*, Shawn Lewis1, Benlian Wang2,3, Masaru Miyagi2,3,4, Puttur Santoshkumar5, Mahesha H. Gangadhariah1 and Ram H. Nagaraj1,3,4,{dagger}

1Department of Ophthalmology & Department of Visual Sciences; 2Center for Proteomics; 3Visual Sciences Research Center; 4Department of Pharmacology, Case Western Reserve University, Cleveland, OH 44106; and 5Mason Eye Institute, University of Missouri, Columbia, MO 65212, USA

{dagger}To whom correspondence should be addressed. Tel: +1-216-368-2089, Fax: +1-216-368-0743, E-mail: ram.nagaraj{at}case.edu

Received December 22, 2007; Accepted March 3, 2008


  Abstract

{alpha}A-crystallin is abundant in the lens of the eye and acts as a molecular chaperone by preventing aggregation of denaturing proteins. We previously found that chemical modification of the guanidino group of selected arginine residues by a metabolic {alpha}-dicarbonyl compound, methylglyoxal (MGO), makes human {alpha}A-crystallin a better chaperone. Here, we examined how the introduction of additional guanidino groups and modification by MGO influence the structure and chaperone function of {alpha}A-crystallin. {alpha}A-crystallin lysine residues were converted to homoarginine by guanidination with o-methylisourea (OMIU) and then modified with MGO. LC-ESI-mass spectrometry identified homoargpyrimidine and homohydroimidazolone adducts after OMIU and MGO treatment. Treatment with 0.25 M OMIU abolished most of the chaperone function. However, subsequent treatment with 1.0 mM MGO not only restored the chaperone function but increased it by ~40% and ~60% beyond that of unmodified {alpha}A-crystallin, as measured with citrate synthase and insulin aggregation assays, respectively. OMIU treatment reduced the surface hydrophobicity but after MGO treatment, it was ~39% higher than control. FRET analysis revealed that {alpha}A-crystallin subunit exchange rate was markedly retarded by OMIU modification, but was enhanced after MGO modification. These results indicate a pattern of loss and gain of chaperone function within the same protein that is associated with introduction of guanidino groups and their neutralization. These findings support our hypothesis that positively charged guanidino group on arginine residues keeps the chaperone function of {alpha}A-crystallin in check and that a metabolic {alpha}-dicarbonyl compound neutralizes this charge to restore and enhance chaperone function.

Key Words: {alpha}A-crystallin, chaperone, homoarginine, homoargpyrimidine, homohydroimidazolone

Abbreviations: sHsps, small heat shock proteins; DTT, dithiothreitol; CS, citrate synthase; OMIU, o-methylisourea; MGO, methylglyoxal; TNS, 2-(p-toludino) naphthalene-6-sulphonic acid, sodium salt; FRET, fluorescence resonance energy transfer; LC-ESI-MS, liquid chromatography-electrospray ionization mass spectrometry; AIAS, 4-acetamido-4'-((iodoacetyl)amino)-stilbene-2,2'-disulophonic acid; LYI, lucifer yellow iodoacetamide

*Present address: Department of Pathobiology, Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.