Frank Jordan
Professor

Rutgers University
Chemistry Department
210 Olson Hall. 73 Warren St.
Newark. NJ 07102
(201) 648-5470
FAX - 1264
frjordan@andromeda.rutgers.edu

Bioorganic and biophysical chemistry. enzymology. protein folding

Over the past decade. research in my lab has focused on the structure. folding. inhibitor design and mechanism of action of two classes of enzymes: thiamin diphosphate dependent a-keto acid decarboxylases and dehydrogenases and the serine proteases. Research on thiamin enzymes includes development of mechanism-based inactivators. and elucidation of active center structure through high resolution x-ray structure determination of brewer's yeast pyruvate decarboxylase. elucidation of the regulation of such enzymes by a combined use of structural and molecular genetic probes. and spectroscopic studies of enzyme-bound intermediates (ranging from UV-vis stopped-flow to NMR). Concurrently with the enzyme studies. we are also synthesizing "bis-coenzyme" models for two oxidative decarboxylases pyruvate oxidase and pyruvate dehydrogenase. By the combined use of high resolution information on the enzymes and appropriate chemical model reactions we are hoping to learn which steps in the complex mechanism require assistance of the protein over and above the catalytic acceleration afforded by the coenzymes. as well as the magnitude of the rate of acceleration that is provided by the protein. and the amino acid side chain(s) responsible for it. Research on serine proteases has included: a) design and synthesis of novel electrophilic groups for specific inhibition; b) multinuclear NMR studies of the active center electronic structure in native and inhibited enzymes and of the ubiquitous Ca(II) sites and c) elucidation of the pro-peptide assisted folding of subtilisins. Having available the genetic tools. recent results on the last problem mentioned are beginning to provide very detailed information about a rather new pathway for protein folding (pioneered by M. Inouye in the late 80's) and this problem is also well suited for high resolution spectroscopic investigations.

Selected Publications

Jordan F. (2007) Adenosine triphosphate and thiamine cross paths. Nat Chem Biol. 3(4):202-3.

Schweitzer-Stenner R, Measey T, Kakalis L, Jordan F, Pizzanelli S, Forte C, Griebenow K. (2007) Conformations of alanine-based peptides in water probed by FTIR, Raman, vibrational circular dichroism, electronic circular dichroism, and NMR spectroscopy. Biochemistry. F46(6):1587-96.

Nemeria N, Chakraborty S, Baykal A, Korotchkina LG, Patel MS, Jordan F. (2007) The 1',4'-iminopyrimidine tautomer of thiamin diphosphate is poised for catalysis in asymmetric active centers on enzymes. Proc Natl Acad Sci U S A. 104(1):78-82.

Joseph E, Wei W, Tittmann K, Jordan F. (2006) Function of a conserved loop of the beta-domain, not involved in thiamin diphosphate binding, in catalysis and substrate activation in yeast pyruvate decarboxylase. Biochemistry. 45(45):13517-27.

Baykal AT. Kakalis L. Jordan F. (2006) Electronic and nuclear magnetic resonance spectroscopic features of the 1',4'-iminopyrimidine tautomeric form of thiamin diphosphate. a novel intermediate on enzymes requiring this coenzyme. Biochemistry. 45(24):7522-8.

Arjunan P. Sax M. Brunskill A. Chandrasekhar K. Nemeria N. Zhang S. Jordan F. Furey W. (2006) A thiamin-bound. pre-decarboxylation reaction intermediate analogue in the pyruvate dehydrogenase E1 subunit induces large scale disorder-to-order transformations in the enzyme and reveals novel structural features in the covalently bound adduct. J Biol Chem. 281(22):15296-303.

Jordan F. Nemeria NS. Sergienko E. (2005) Multiple modes of active center communication in thiamin diphosphate-dependent enzymes. Acc Chem Res. 38(9):755-63.

Jordan F. Nemeria NS. (2005) Experimental observation of thiamin diphosphate-bound intermediates on enzymes and mechanistic information derived from these observations. Bioorg Chem. 33(3):190-215.

Nemeria N. Tittmann K. Joseph E. Zhou L. Vazquez-Coll MB. Arjunan P. Hubner G. Furey W. Jordan F.
(2005) Glutamate 636 of the Escherichia coli pyruvate dehydrogenase-E1 participates in active center communication and behaves as an engineered acetolactate synthase with unusual stereoselectivity. J Biol Chem. 280(22):21473-82.

Zhang S. Zhou L. Nemeria N. Yan Y. Zhang Z. Zou Y. Jordan F. (2005) Evidence for dramatic acceleration of a C-H bond ionization rate in thiamin diphosphate enzymes by the protein environment. Biochemistry. 44(7):2237-43.

Park YH. Wei W. Zhou L. Nemeria N. Jordan F. (2004) Amino-terminal residues 1-45 of the Escherichia coli pyruvate dehydrogenase complex E1 subunit interact with the E2 subunit and are required for activity of the complex but not for reductive acetylation of the E2 subunit. Biochemistry. 43(44):14037-46.

Jordan F. (2004) Biochemistry. How active sites communicate in thiamine enzymes. Science. 306(5697):818-20.

Zhang S. Liu M. Yan Y. Zhang Z. Jordan F. (2004) C2-alpha-lactylthiamin diphosphate is an intermediate on the pathway of thiamin diphosphate-dependent pyruvate decarboxylation. Evidence on enzymes and models. J Biol Chem. 279(52):54312-8.

Nemeria N. Baykal A. Joseph E. Zhang S. Yan Y. Furey W. Jordan F. (2004) Tetrahedral intermediates in thiamin diphosphate-dependent decarboxylations exist as a 1',4'-imino tautomeric form of the coenzyme. unlike the michaelis complex or the free coenzyme. Biochemistry. 43(21):6565-75.

Arjunan P. Chandrasekhar K. Sax M. Brunskill A. Nemeria N. Jordan F. Furey W. (2004) Structural determinants of enzyme binding affinity: the E1 component of pyruvate dehydrogenase from Escherichia coli in complex with the inhibitor thiamin thiazolone diphosphate. Biochemistry. 43(9):2405-11.

Jordan F. Nemeria NS. Zhang S. Yan Y. Arjunan P. Furey W. (2003) Dual catalytic apparatus of the thiamin diphosphate coenzyme: acid-base via the 1',4'-iminopyrimidine tautomer along with its electrophilic role. J Am Chem Soc. 125(42):12732-8.

Tittmann K. Golbik R. Uhlemann K. Khailova L. Schneider G. Patel M. Jordan F. Chipman DM. Duggleby RG. Hubner G. (2003) NMR analysis of covalent intermediates in thiamin diphosphate enzymes. Biochemistry. 42(26):7885-91.

Jordan F. (2003) Current mechanistic understanding of thiamin diphosphate-dependent enzymatic reactions. Nat Prod Rep. 20(2):184-201.

Wei W. Li H. Nemeria N. Jordan F. (2003) Expression and purification of the dihydrolipoamide acetyltransferase and dihydrolipoamide dehydrogenase subunits of the Escherichia coli pyruvate dehydrogenase multienzyme complex: a mass spectrometric assay for reductive acetylation of dihydrolipoamide acetyltransferase. Protein Expr Purif. 28(1):140-50.

Polovnikova ES. McLeish MJ. Sergienko EA. Burgner JT. Anderson NL. Bera AK. Jordan F. Kenyon GL. Hasson MS. (2003) Structural and kinetic analysis of catalysis by a thiamin diphosphate-dependent enzyme. benzoylformate decarboxylase. Biochemistry. 42(7):1820-30.

Arjunan. P.. Nemeria. N.. Brunskill. A.. Chandrasekhar. K.. Sax. M.. Yan. Y.. Jordan. F.. Guest J.R. and Furey. W. (2002)Structure of the pyruvate dehydrogenase multienzyme complex E1 Component from E. coli at 1.85 Ċ resolution. Biochemistry. 41(16):5213-21.

Sergienko. E.A.. and Jordan. F. (2002) Yeast pyruvate decarboxylase tetramers can dissociate into dimers along two interfaces. Hybrids of low-activity D28A (or D28N) and E477Q variants. with substitution of adjacent active center acidic groups from different subunits. display restored activity. Biochemistry. 41(19):6164-9.

Wei. W.. Min Liu. M. and Jordan. F. (2002) Solvent kinetic isotope effects monitor changes in hydrogen bonding at the active center of yeast pyruvate decarboxylase concomitant with substrate activation. The substituent at Position 221 can control the state of activation. Biochemistry 41:451-461.

Kahyaoglu. A. and Jordan. F. (2002) Direct proton magnetic resonance determination of the pKa of the active center histidine in thiolsubtilisin. Protein Science 11:965-973.

Sergienko E. A. and Jordan. F. (2002) A new model for activation of yeast pyruvate decarboxylase by substrate consistent with the alternating sites mechanism: Demonstration of the existence of two active forms of the enzyme. Biochemistry 41:3952-3967.

Wang. J.. Golbik. R... Seliger. B.. Spinka. M.. Tittmann. K.. Hübner. G.. and Jordan. F. (2001) Consequences of a modified putative substrate-activation site on catalysis by yeast pyruvate decarboxylase. Biochemistry 40:1755-1763.

Liu. M.. Eduard A. Sergienko. E.A.. Guo. F.. Wang. J.. Tittmann. K.. Hübner. G.. Furey. W. and Jordan. F. (2001) Catalytic acid-base groups in yeast pyruvate decarboxylase I. Site directed mutagenesis and steady-state kinetic studies on the enzyme with the D28A. H114F. H115F and E477Q substitutions. Biochemistry 40:7355-7368.

Sergienko. E. A.. and Jordan. F. (2001) Catalytic acid-base groups in yeast pyruvate decarboxylase Ii. Insights to the specific roles of D28 and E477 from the rates and stereospecificity of formation of carboligase side products. Biochemistry 40:7369-7381.

Sergienko. E. A.. and Jordan. F. (2001) Catalytic acid-base groups in yeast pyruvate decarboxylase Iii. A steady-state kinetic model consistent with the behavior of both wild-type and variant enzymes at all relevant pH values. Biochemistry 40:7382-7403.

Nemeria. N.. Yan. Y.. Zhang. Z.. Brown. A.M.. Arjunan. P.. Furey. W.. Guest. J.R. and Jordan. F. (2001) Inhibition of the E. coli pyruvate dehydrogenase complex-E1 subunit and its Tyrosine 177 variants by Thiamin 2-Thiazolone and Thiamin 2-Thiothiazolone Diphosphates: evidence for reversible tight-binding inhibition. J. Biol. Chem. 49:45969-45978.