U.I.C. Biochemistry - Dr. Gettins

Dr. Peter G.W. Gettins


Director, Center for Structural Biology

D. Phil,1979, Oxford University

Postdoctoral, Yale

MAJOR INTERESTS: Structure and function of the serpin superfamily. Antithrombin structure, function and dysfunction. Structure, specificity and function of the LDL receptor-related protein LRP and its protein ligands. Use of NMR and other spectroscopic methods, as well as kinetic and thermodynamic approaches to determine structure/function correlations in solution. Use of x-ray crystallography as complement to solution studies.

1.SERPINS. Serpins are large proteins (45-100kDa) that act as inhibitors, and thus regulators, of important physiological processes such as blood coagulation and fibrinolysis, and that employ a remarkable conformational change mechanism for inhibition of target proteinases. Serpins being currently studied are antithrombin, alpha-1-antitrypsin (alpha-1-proteinase inhibitor),crmA, pigment epithelium derived factor (PEDF) and heparin cofactor II. Antithrombin is the principal inhibitor of the blood coagulation proteinases thrombin and factor Xa; alpha-1-antitrypsin is the principal inhibitor of neutrophil elastase; crmA (cytokine response modifier A) is a viral serpin that the virus uses to arrest cell death of the host by inhibition of caspases and granzyme B proteinases that normally orchestrate cell death; pigment epithelium derived factor is a non-inhibitory member of the serpin superfamily that has potent anti-angiogenic activity and neurite outgrowth-promoting activity; heparin cofactor II is another plasma inhibitor of thrombinthat is activated by various glycosaminoglycans.

Questions being addressed are

Methods being used are

2. LRP. LRP is a 600kDa multidomain receptor of the LDL receptor family. It binds and clears from circulation an extremely wide range of unrelated protein ligands, including serpin-proteinase complexes, the 720kDa non-specific proteinase inhibitor alpha-2-macroglobulin when complexed with proteinases, certain lipoproteins,the 39kDa chaperone protein RAP( receptor associated protein) as well as a number of other ligands. We are interested in determining the structural basis for the ability of LRP to bind to such a wide range of unrelated proteins. To address this we are detemining the structure of domains of LRP that bind ligands and of complexes of these domains, whether single or larger species, in complex with target protein ligands .


Recent Publications

Graduate and postdoctoral students are indicated in bold type

Qazi, U., Gettins, P.G.W., Strickland, D.K., and Stoops, J.K. (1999) J. Biol. Chem. 274, 8137-8142. "Structural details of proteinase entrapment by human alpha-2-macroglobulin emerge from three-dimensional reconstructions of Fab-labeled native, half-transformed and transformed molecules."

Stratikos, E., and Gettins, P.G.W. (1999) Proc. Natl. Acad. Sci. USA 96, 4803-4813. "Formation of the covalent serpin-proteinase complex involves translocation of the proteinase by more than 70 Å and full insertion of the reactive center loop into beta-sheet A.

Huang, W., Dolmer, K., and Gettins, P.G.W. (1999) J. Biol. Chem. 274, 14130-14136. "NMR solution structure of complement-like repeat CR8 from the low density lipoprotein receptor-related protein (LRP)."

Chuang, Y.-J., Gettins, P.G.W., and Olson, S.T. (1999) J. Biol. Chem. 274, 28142-28149. "Importance of the P2 glycine of antithrombin in target proteinase specificity, heparin activation, and the efficiency of proteinase trapping as revealed by a P2 Gly-to-Pro mutation."

Huang, W., Dolmer, K., Liao, X., and Gettins, P.G.W. (2000) J. Biol. Chem. 275, 1089-1094. "NMR solution structure of the receptor binding domain of human alpha-2-macroglobulin."

Meagher, J.L.,, Olson, S.T., and Gettins, P.G.W. (2000) J. Biol. Chem. 275, 2698-2704. "Critical role of the linker region between helix D and strand 2A in heparin activation of antithrombin."

Dolmer, K., Huang, W., and Gettins, P.G.W. (2000) J. Biol. Chem. 275, 3264-3271. "NMR solution structure of complement-like repeat CR3 from the low density lipoprotein-receptor-related protein (LRP). Evidence for specific binding to the receptor binding domain of human alpha-2-macroglobulin."

Futamura, A.,, and Gettins, P.G.W. (2000) J. Biol. Chem. 275, 4092-4098. "Serine 380 (P14) to glutamate mutation activates antithrombin as an inhibitor of factor Xa."

Huntington, J.A., McCoy, A., Pei, X., Gettins, P.G.W., and Carrell, R.W. (2000) J.Biol.Chem. 275, 15377-15383. "The conformational activation of antithrombin. A 2.85 Å structure of fluorescein derivative reveals an electrostatic linkage between the hinge and heparin binding regions."

Qazi, U., Kolodziej, S.J., Gettins, P.G.W., and Stoops, J.K. (2000) J. Struct. Biol. 131, 19-26. The structure of the C949S mutant alpha-2-macroglobulin demonstrates the role of the internal thiol esters in proteinase-entrapping structural transformation."

Simonovic,M., Gettins, P.G.W., and Volz, K. (2000) Acta Crystallographica D56, 1440-1442. "Crystallization and preliminary x-ray analysis of recombinant cysteine-free variant of CrmA."

Simonovic, M., Gettins, P.G.W., and Volz, K. (2000) Protein Science 9, 1423-1427. Accelerated Communication: "Crystal structure of viral serpin CrmA provides insights into its mechanism of cysteine proteinase inhibition."

Sivasothy, P., Dafforn, T.R., Gettins, P.G.W., and Lomas, D.A. (2000) J. Biol. Chem. 275, 33663-33668. "Pathogenic alpha-1-antitrypsin polymers are formed by reactive loop-beta-sheet A linkage."

Peterson, F., Gordon, N., and Gettins, P.G.W. (2000) Biochemistry 39, 11884-11892. "Formation of a non-covalent serpin-proteinase complex involves no conformational change in the serpin. Use of 1H-15N HSQC NMR as a sensitive non-perturbing monitor of conformation."

Suda, S.A., Gettins, P.G.W., and Patston, P.A. (2000) Arch. Biochem. Biophys. 384, 31-36. "Linkage between the hormone binding site and reactive center loop of thyroxine binding globulin."

Gettins, P.G.W. (2000) Genome Research, 10, 1833-1835. "Keeping the serpin machine running smoothly." Invited commentary.

Wang, Q.-Y., Dolmer, K., Huang, W., Gettins, P.G.W., and Rong, L. (2001) J. Virol. 75, 2051-2058. "Role of calcium in protein folding and function of Tva, the receptor of subgroup A avian sarcoma and leukosis virus"

Peterson, F.C., Gordon, N., and Gettins, P.G.W. (2001) Biochemistry 40, 6275-6283. "High level bacterial expression and 15N-alanine labelling of bovine trypsin. Application to study of trypsin-inhibitor complexes and trypsinogen activation by NMR spectroscopy"

Peterson, F.C. and Gettins, P.G.W. (2001) Biochemistry 40, 6284-6292. "Insight into the mechanism of serpin-proteinase inhibition from 2D [1H-15N] NMR studies of the 69kDa alpha-1-proteinase inhibitor Pittsburgh-trypsin covalent complex"

Futamura, A., Beechem, J., and Gettins, P.G.W. (2001) Biochemistry 40, 6680-6687. "Conformational equilibrium of the reactive center loop of antithrombin: Examination by steady state and time-resolved fluorescence".

Simonovic. M., Gettins, P.G.W., and Volz, K. (2001) Proc. Natl. Acad. Sci. USA,98, 11131-11135. "Crystal structure of human PEDF, a potent anti-angiogenic and neurite outgrowth-promoting factor"

Simonovic, M., Dolmer, K., Huang, W., Strickland, D.K., Volz, K., and Gettins, P.G.W. (2001) Biochemistry 40, 15127-15134. "X-ray structure and calcium affinity of ligand-binding repeat CR7 from LRP. Comparison with related domains from LRP and the LDL receptor."

Silverman, G.A., Bird, P.I., Carrell, R.W., Church, F.C., Coughlin, P.B., Gettins, P.G.W., Irving, J., Lomas, D.A., Moyer, R.W., Pemberton, P., Remold-O'Donnell, E., Salvesen, G., Travis, J., and Whisstock, J. (2001) J. Biol. Chem. 276, 33293-33296. "The serpins are an expanding superfamily of structurally similar, but functionally diverse proteins."

Wang, Q.-Y., Huang, W., Dolmer, K., Gettins, P.G.W., and Rong, L. (2002) J. Virol. 76, 2848-2856. "NMR solution structure of the viral receptor domain of Tva and its implications in viral entry."

Belzar, K.J., Zhou, A., Carrell, R.W., Gettins, P.G.W., and Huntington, J.A. (2002) J. Biol. Chem. 277, 8551-8558. "Helix D elongation and allosteric activation of antithrombin."

Backovic, M., Stratikos, E., Lawrence, D.A., and Gettins, P.G.W. (2002) Prot. Sci. 11, 1182-1191. Structural similarity of the covalent complexes formed between the serpin plasminogen activator inhibitor-1 and the arginine-specific proteinases trypsin, LMW urokinase, HMW urokinase and t-PA: Use of site-specific fluorescent probes of local environment."

Schedin-Weiss, S., Desai, U.R., Bock, S.C., Gettins, P.G.W., Olson, S.T., and Björk, I. (2002) Biochemistry, 41, 4770-4788. "The importance of lysine 125 for heparin binding and activation of antithrombin."

Backovic, M., and Gettins, P.G.W. (2002) J. Proteome Res. 1, 367-373. "Insight into residues critical for antithrombin function from an expanded database that includes sequences of frog, turtle and ostrich antithrombins."

Gettins, P.G.W. (2002) FEBS Lett. 523, 2-6. "Helix F plays an active, critical role in the serpin mechanism."

Gettins, P.G.W. (2002) Chem. Rev. 102, 4751-4804. "Serpin structure, function and biology. "

Gettins, P.G.W., Simonovic, M., and Volz, K. (2002) Biol. Chem. 383, 1677-1682. "Pigment epithelium-derived factor (PEDF), a serpin with potent anti-angiogenic and neurite outgrowth-promoting properties."

Wang, Q.-Y., Manicassamy, B., Yu, X., Dolmer, K., Gettins, P.G.W. and Rong, L. (2002) Prot. Sci. 11, 2596-2605. "Characterization of the LDL-A module mutants of Tva, the subgroup A Rous sarcoma virus receptor, and the implications for protein folding."

Yu, X., Wang, Q.-Y., Guo, Y., Dolmer, K., Young, J.A., Gettins, P.G.W., and Rong, L. (2003) J. Virol. 77, 7517-7526. "Kinetic analysis of binding interaction between the subgroup A Rous sarcoma virus glycoprotein SU and its cognate receptor Tva: calcium is not required for ligand binding."

Dementiev, A., Simonovic, M., Volz, K., and Gettins, P.G.W. (2003) J. Biol. Chem. ,278, 37881-37887. "Canonical inhibitor-like interactions explain reactivity of alpha-1-proteinase inhibitor Pittsburgh and antithrombin with proteinases."

Lazic, A., Dolmer, K., Strickland, D.K., and Gettins, P.G.W. (2003) Biochemistry, 42, 14913-14920. " Structural organization of the receptor associated protein."

Dobo, J., and Gettins, P.G.W. (2004) J. Biol. Chem., 279, 9264-9269. "Alpha-1-proteinase inhibitor forms initial non-covalent and final covalent complexes with elastase analogously to other serpin-proteinase pairs, suggesting a common mechanism of inhibition."

Dementiev, A., Petitou, M., Herbert, J.M., and Gettins, P.G.W. (2004) Nat. Struct. Mol. Biol. 11, 863-867. "The ternary complex of antithrombin-anhydrothrombin-heparin reveals the basis of inhibitory specificity."

Gettins, P.G.W., and Olson, S.T. (2004) Methods 32, 110-119. "Use of fluorescence resonance energy transfer to study serpin-proteinase interactions."

Gettins, P.G.W., Backovic, M., and Peterson, F.C. (2004) Methods 32, 120-129. "Use of NMR to study serpin function".

Al-Ayyoubi, M., Gettins, P.G.W., and Volz, K. (2004) J. Biol. Chem. 279, 55540-55544. "Crystal structure of human maspin, a serpin with anti-tumor properties: Maspin's reactive center loop is exposed but constrained."

Tesch, L.D., Raghavendra, M.P., Bedsted-Faarvang, T., Gettins, P.G.W., and Olson, S.T. (2005) Prot. Sci. 14, 533-542. "Specificity and reactive loop length requirements for crmA inhibition of serine proteinases."

Gettins, P.G.W., and Dolmer, K. (2005) "Complement-like repeats in proteins of the complement system" in "Structural biology of the complement system" J. Lambris ed. Marcel Dekker, pp265-292

Simonovic, M., Denault, J.-B., Salvesen, G.S., Volz, K., and Gettins, P.G.W. (2005) Arch. Biochem. Biophys. 440, 1-9. "Lack of involvement of strand s1'A of the viral serpin CrmA in anti-apoptotic or caspase-inhibitory functions."

Dementiev, A., Dobo, J., and Gettins, P.G.W. (2006) J. Biol. Chem. 281, 3452-3457. "Active site distortion is sufficient for proteinase inhibition by serpins. Structure of the covalent complex of alpha-1-proteinase inhibitor with porcine pancreatic elastase."

Lazic, A., Dolmer, K., Strickland, D.K., and Gettins, P.G.W. (2006) Arch. Biochem. Biophys. 450, 167-175. ssection of RAP-LRP interactions. Binding of RAP and RAP fragments to complement-like repeats 7 and 8 from ligand binding cluster II of LRP."

Dolmer, K., and Gettins, P.G.W. (2006) J. Biol. Chem. 281, 34189-34196. "Three complement-like repeats compose the complete alpha-2-macroglobulin binding site in the second ligand binding cluster of the low density lipoprotein receptor-related protein."

Dobo, J., Swanson, R., Salvesen, G.S., Olson, S.T., and Gettins, P.G.W. (2006) J. Biol. Chem. 281, 38781-38790. "Cytokine response modifier A inhibition of initiator caspases results in covalent complex formation and dissociation of the caspase tetramer."

Swanson, R., Raghavendra, M.P., Zhang, W., Froelich, C., Gettins, P.G.W. and Olson, S.T., (2007) J. Biol. Chem. 282, 2305-2313. "Serine and cysteine proteases are translocated to similar extents upon formation of covalent complexes with serpins."

Gettins, P.G.W. (2007) "Mechanisms of serpin inhibition", in Molecular and Cellular Aspects of the Serpinopathies and Disorders in Serpin Activity, G.S. Silverman, ed. in press.

Current members of the lab

Recent past members of the lab

send mail to pgettins@uic.edu

last updated 24th January, 2007