HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sharma, S. Articles by Waygood, E. B. Search for Related Content PubMed PubMed Citation Articles by Sharma, S. Articles by Waygood, E. B. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 268, Issue 24, 17695-17704, 08, 1993

Deamidation of HPr, a phosphocarrier protein of the phosphoenolpyruvate:sugar phosphotransferase system, involves asparagine 38 (HPr-1) and asparagine 12 (HPr-2) in isoaspartyl acid formation

S Sharma, PK Hammen, JW Anderson, A Leung, F Georges, W Hengstenberg, RE Klevit and EB Waygood
Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

Histidine-containing protein, HPr, of the phosphoenolpyruvate:sugar phosphotransferase system in Escherichia coli, when incubated at elevated temperatures forms many species of protein. The two major species are HPr-1 and HPr-2, which have been shown to lack one or two amides, respectively (Anderson, B., Weigel, N., Kundig, W., and Roseman, S. (1971) J. Biol. Chem. 246, 7023-7033). The formation of HPr- 1 and HPr-2 is shown to be pH-dependent and does not occur readily below pH 6. Investigation of the identities and properties of the two residues that deamidate involved creation of site-directed mutants at the 6 glutamine and 2 asparagine residues of HPr; description of their deamidation species by isoelectric focusing; determination of their relative antibody binding properties; assay of their phosphoacceptor and phosphodonor activities; characterization of tryptic and V8- protease peptides; obtaining two-dimensional nuclear magnetic resonance spectra of HPr, HPr-1, and several mutants. It was determined that the sequential deamidation of Asn-38 and Asn-12 yields HPr-1 and HPr-2. Both residues exist as Asn-Gly pairs, and both deamidations probably form isoaspartyl acid. HPr from Bacillus subtilis and Staphylococcus carnosus which also have Asn-Gly at residues 38 and 39 form HPr-1 species presumably by deamidation. HPr from Streptococcus faecalis which does not have Asn-38 does not form a HPr-1 species. The E. coli mutant HPrs, N12D and Q51E, residues that may be involved in the active site, had impaired phosphohydrolysis properties and decreased phosphoenolpyruvate:sugar phosphotransferase system activity.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Kindrachuk, J. Parent, G. F. Davies, M. Dinsmore, S. Attah-Poku, and S. Napper Overexpression of L-Isoaspartate O-Methyltransferase in Escherichia coli Increases Heat Shock Survival by a Mechanism Independent of Methyltransferase Activity J. Biol. Chem., December 19, 2003; 278(51): 50880 - 50886. [Abstract] [Full Text] [PDF]

				L. Athmer, J. Kindrachuk, F. Georges, and S. Napper The Influence of Protein Structure on the Products Emerging from Succinimide Hydrolysis J. Biol. Chem., August 16, 2002; 277(34): 30502 - 30507. [Abstract] [Full Text] [PDF]

				N. E. Robinson and A. B. Robinson Prediction of protein deamidation rates from primary and three-dimensional structure PNAS, April 10, 2001; 98(8): 4367 - 4372. [Abstract] [Full Text] [PDF]

				M. J. Mamula, R. J. Gee, J. I. Elliott, A. Sette, S. Southwood, P.-J. Jones, and P. R. Blier Isoaspartyl Post-translational Modification Triggers Autoimmune Responses to Self-proteins J. Biol. Chem., August 6, 1999; 274(32): 22321 - 22327. [Abstract] [Full Text] [PDF]

				S. Napper, L. T. J. Delbaere, and E. B. Waygood The Aspartyl Replacement of the Active Site Histidine in Histidine-containing Protein, HPr, of the Escherichia coli Phosphoenolpyruvate:Sugar Phosphotransferase System Can Accept and Donate a Phosphoryl Group. SPONTANEOUS DEPHOSPHORYLATION OF ACYL-PHOSPHATE AUTOCATALYZES AN INTERNAL CYCLIZATION J. Biol. Chem., July 30, 1999; 274(31): 21776 - 21782. [Abstract] [Full Text] [PDF]