PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• Contributors to Volume 343.Preface.Volume in Series.Section I. G Protein-Coupled Receptors.A. Theoretical Evaluation of Receptor Function:Considerations in the Evaluation of Inverse Agonism and Protean Agonism at G Protein-Coupled Receptors,P. Chidiac.Theoretical Implications of Receptor Coupling to Multiple G Proteins Based on Analysis of a Three-State Model,C. Scaramellini and P. Leff.B. Design and Use of Receptor Ligands:Use of Retinal Analogues for the Study of Visual Pigment Function,R.K. Crouch, V. Kefalov, W. Gartner, and M.C. Cornwall.Design and Synthesis of Peptide Antagonists and Inverse Agonists for G Protein-Coupled Receptors,S.M. Cowell, P.M. Balse-Srinivasan, J.M. Ahn, and V.J. Hruby.Design of Peptide Agonists,V.J. Hruby, R.S. Agnes, C. Cai.Design of Nonpeptides from Peptide Ligands for Peptide Receptors,V.J. Hruby, W. Qiu, T. Okayama, and V.A. Soloshonok.Strategies for Mapping the Binding Site of the Serotonin 5-HT2A Receptor,B.J. Ebersole and S.C. Sealfon.C. Structural Characterization of Receptor Proteins:Use of the Substituted Cysteine Accessibility Method to Study the Structure and Function of G Protein-Coupled Receptors,J.A. Javitch, L. Shi, and G. Liapakis.Mass Spectrometric Analysis of G Protein-Coupled Receptors,D.R. Knapp, R.K. Crouch, L.E. Ball, A.K. Gelasco, and Z. Ablonczy.Probing the Higher Order Structure of G protein-Coupled Receptors Using Tethered Cleavage Methods,A.K. Gelasco, R.K. Crouch, and D.R. Knapp.Use of Fluorescence Spectroscopy to Study Conformational Changes in the b2-Adrenoceptor,B.K. Kobilka and U. Gether.Crystallization of Membrane Protein in Cubo,P. Nollert, J. Navarro, and E.M. Landau.N-Linked Carbohydrates on G Protein-Coupled Receptors: Mapping Sites of Attachment and Determining Functional Roles,D.P. Davis and D.L. Segaloff.Magic Angle Spinning Nuclear Magnetic Resonance of Isotropically Labeled Rhodopsin,M. Eilers, W. Ying, P.J. Reeves, H.G. Khorana, and S.O. Smith.Use of Nuclear Magnetic Resonance to Study the Three-Dimensional Structure of Rhodopsin,P.L. Yeagle and A.D. Albert.D. Design and Use of Engineered Receptor Proteins:Tools for Dissecting Signaling Pathways in Vivo: Receptors Activated Solely by Synthetic Ligands,K. Scearce-Levie, P. Coward, C.H. Redfern, and B.R. Conklin.Analysis of Structure-Function from Expression of G Protein-Coupled Receptor Fragments,S.S. Karnik.Construction and Analysis of Function of G Protein-Coupled Receptor-G Protein Fusion Proteins,G. Milligan.Synthetic Gene Technology: Applications to Ancestral Gene Reconstruction and Structure-Function Studies of Receptors,B.S.W. Chang, M.A. Kazmi, and T.P. Kakmar.Considerations in the Design and Use of Chimeric G Protein-Coupled Receptor,J. Wess.E. Molecular Modeling Studies of Receptor Structure and Function:Strategies for Modeling the Interactions of Transmembrane Helices of G Protein-Coupled Receptors by Geometric Complementarity Using the GRAMM Computer Algorithm,I.A. Vakser and S. Jiang.Three-Dimensional Representation of G Protein-Coupled Receptor Structures and Mechanisms,I. Visiers, J.A. Ballesteros, and H. Weinstein.F. Analysis of Receptor Protein Coupling:Reconstitution of G Protein-Coupled Receptors with Recombinant G Protein a and bg Subunits,W.E. McIntire, C.S. Myung, G. MacCleery, Q. Wang, and J.C. Garrison.Cell-Free Membrane Desensitization Assay for G Protein-Coupled Receptors,M. Hunzicker-Dunn and L. Birnbaumer.Methods to Determine the Constitutive Activity of Histamine H2 Receptors,R. Leurs, M. Hoffman, A.E. Alewijnse, M.J. Smit, and H. Timmerman.Expression of G Protein-Coupled Receptors and G Proteins in Sf9 Cells: Analysis of Coupling by Radioligand Binding,R.T. Windh and D.R. Manning.G Protein-Coupled Receptors and Proliferative Signaling,M.J. Smit, R.A. Bakker, and E.S. Burstein.G. Characterization of Receptor Heterogeneity:Genetic Analysis of G Protein-Coupled Receptor Genes,D.P. Healy.Identification of Adrenergic Receptor Polymorphisms,K.M. Small, D.A. Rathz, and S.B. Liggett.Strategies and Requirements for the Detection of RNA Editing in G Protein Coupled-Receptor RNA,C.M. Niswender.H. The Study of Receptor Trafficking:Fluorescence Microscopy Techniques for the Study of G Protein-Coupled Receptor Trafficking,L. Kallal and J.L. Benovic.Measurement of Receptor Desensitization and Internalization in Interact Cells,R.B. Clark and B.J. Knoll.Morphological and Biochemical Strategies for Monitoring Trafficking of Epitope-Tagged G Protein-Coupled Receptors in Agonist-Naïve and Agonist-Occupied States,M. Wozniak, C. Saunders, N. Schramm, J.R. Keefer, and L.E. Limbird.Section II. Regulators of GPCR Function.A. G Protein-Coupled Receptor Kinases (GRKs):Characterization of G Protein-Coupled Receptor Kinases,A.N. Pronin, R.P. Loudon, and J.L. Benovic.Regulation of G Protein-Coupled Receptor Kinase 2,T. Haga, K. Haga, K. Kameyama, H. Tsuga, and N. Yoshida.Rhodopsin and Its Kinase,I. Sokal, A. Pulvermuller, J. Buczylko, K.P. Hofmann, and K. Palczewski.B. Arrestins and Novel Proteins:Characterization of Arrestin Expression and Function,S.J. Mundell, M.J. Orsini, and J.L. Benovic.Identification of Novel G Protein-Coupled Receptor-Interacting Proteins,R.T. Premont and R.A. Hall.Author Index.Subject Index.

PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• Contributors to Volume 343.Preface.Volume in Series.Section I. G Protein-Coupled Receptors.A. Theoretical Evaluation of Receptor Function:Considerations in the Evaluation of Inverse Agonism and Protean Agonism at G Protein-Coupled Receptors,P. Chidiac.Theoretical Implications of Receptor Coupling to Multiple G Proteins Based on Analysis of a Three-State Model,C. Scaramellini and P. Leff.B. Design and Use of Receptor Ligands:Use of Retinal Analogues for the Study of Visual Pigment Function,R.K. Crouch, V. Kefalov, W. Gartner, and M.C. Cornwall.Design and Synthesis of Peptide Antagonists and Inverse Agonists for G Protein-Coupled Receptors,S.M. Cowell, P.M. Balse-Srinivasan, J.M. Ahn, and V.J. Hruby.Design of Peptide Agonists,V.J. Hruby, R.S. Agnes, C. Cai.Design of Nonpeptides from Peptide Ligands for Peptide Receptors,V.J. Hruby, W. Qiu, T. Okayama, and V.A. Soloshonok.Strategies for Mapping the Binding Site of the Serotonin 5-HT2A Receptor,B.J. Ebersole and S.C. Sealfon.C. Structural Characterization of Receptor Proteins:Use of the Substituted Cysteine Accessibility Method to Study the Structure and Function of G Protein-Coupled Receptors,J.A. Javitch, L. Shi, and G. Liapakis.Mass Spectrometric Analysis of G Protein-Coupled Receptors,D.R. Knapp, R.K. Crouch, L.E. Ball, A.K. Gelasco, and Z. Ablonczy.Probing the Higher Order Structure of G protein-Coupled Receptors Using Tethered Cleavage Methods,A.K. Gelasco, R.K. Crouch, and D.R. Knapp.Use of Fluorescence Spectroscopy to Study Conformational Changes in the b2-Adrenoceptor,B.K. Kobilka and U. Gether.Crystallization of Membrane Protein in Cubo,P. Nollert, J. Navarro, and E.M. Landau.N-Linked Carbohydrates on G Protein-Coupled Receptors: Mapping Sites of Attachment and Determining Functional Roles,D.P. Davis and D.L. Segaloff.Magic Angle Spinning Nuclear Magnetic Resonance of Isotropically Labeled Rhodopsin,M. Eilers, W. Ying, P.J. Reeves, H.G. Khorana, and S.O. Smith.Use of Nuclear Magnetic Resonance to Study the Three-Dimensional Structure of Rhodopsin,P.L. Yeagle and A.D. Albert.D. Design and Use of Engineered Receptor Proteins:Tools for Dissecting Signaling Pathways in Vivo: Receptors Activated Solely by Synthetic Ligands,K. Scearce-Levie, P. Coward, C.H. Redfern, and B.R. Conklin.Analysis of Structure-Function from Expression of G Protein-Coupled Receptor Fragments,S.S. Karnik.Construction and Analysis of Function of G Protein-Coupled Receptor-G Protein Fusion Proteins,G. Milligan.Synthetic Gene Technology: Applications to Ancestral Gene Reconstruction and Structure-Function Studies of Receptors,B.S.W. Chang, M.A. Kazmi, and T.P. Kakmar.Considerations in the Design and Use of Chimeric G Protein-Coupled Receptor,J. Wess.E. Molecular Modeling Studies of Receptor Structure and Function:Strategies for Modeling the Interactions of Transmembrane Helices of G Protein-Coupled Receptors by Geometric Complementarity Using the GRAMM Computer Algorithm,I.A. Vakser and S. Jiang.Three-Dimensional Representation of G Protein-Coupled Receptor Structures and Mechanisms,I. Visiers, J.A. Ballesteros, and H. Weinstein.F. Analysis of Receptor Protein Coupling:Reconstitution of G Protein-Coupled Receptors with Recombinant G Protein a and bg Subunits,W.E. McIntire, C.S. Myung, G. MacCleery, Q. Wang, and J.C. Garrison.Cell-Free Membrane Desensitization Assay for G Protein-Coupled Receptors,M. Hunzicker-Dunn and L. Birnbaumer.Methods to Determine the Constitutive Activity of Histamine H2 Receptors,R. Leurs, M. Hoffman, A.E. Alewijnse, M.J. Smit, and H. Timmerman.Expression of G Protein-Coupled Receptors and G Proteins in Sf9 Cells: Analysis of Coupling by Radioligand Binding,R.T. Windh and D.R. Manning.G Protein-Coupled Receptors and Proliferative Signaling,M.J. Smit, R.A. Bakker, and E.S. Burstein.G. Characterization of Receptor Heterogeneity:Genetic Analysis of G Protein-Coupled Receptor Genes,D.P. Healy.Identification of Adrenergic Receptor Polymorphisms,K.M. Small, D.A. Rathz, and S.B. Liggett.Strategies and Requirements for the Detection of RNA Editing in G Protein Coupled-Receptor RNA,C.M. Niswender.H. The Study of Receptor Trafficking:Fluorescence Microscopy Techniques for the Study of G Protein-Coupled Receptor Trafficking,L. Kallal and J.L. Benovic.Measurement of Receptor Desensitization and Internalization in Interact Cells,R.B. Clark and B.J. Knoll.Morphological and Biochemical Strategies for Monitoring Trafficking of Epitope-Tagged G Protein-Coupled Receptors in Agonist-Naïve and Agonist-Occupied States,M. Wozniak, C. Saunders, N. Schramm, J.R. Keefer, and L.E. Limbird.Section II. Regulators of GPCR Function.A. G Protein-Coupled Receptor Kinases (GRKs):Characterization of G Protein-Coupled Receptor Kinases,A.N. Pronin, R.P. Loudon, and J.L. Benovic.Regulation of G Protein-Coupled Receptor Kinase 2,T. Haga, K. Haga, K. Kameyama, H. Tsuga, and N. Yoshida.Rhodopsin and Its Kinase,I. Sokal, A. Pulvermuller, J. Buczylko, K.P. Hofmann, and K. Palczewski.B. Arrestins and Novel Proteins:Characterization of Arrestin Expression and Function,S.J. Mundell, M.J. Orsini, and J.L. Benovic.Identification of Novel G Protein-Coupled Receptor-Interacting Proteins,R.T. Premont and R.A. Hall.Author Index.Subject Index.

PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• Contributors to Volume 343.Preface.Volume in Series.Section I. G Protein-Coupled Receptors.A. Theoretical Evaluation of Receptor Function:Considerations in the Evaluation of Inverse Agonism and Protean Agonism at G Protein-Coupled Receptors,P. Chidiac.Theoretical Implications of Receptor Coupling to Multiple G Proteins Based on Analysis of a Three-State Model,C. Scaramellini and P. Leff.B. Design and Use of Receptor Ligands:Use of Retinal Analogues for the Study of Visual Pigment Function,R.K. Crouch, V. Kefalov, W. Gartner, and M.C. Cornwall.Design and Synthesis of Peptide Antagonists and Inverse Agonists for G Protein-Coupled Receptors,S.M. Cowell, P.M. Balse-Srinivasan, J.M. Ahn, and V.J. Hruby.Design of Peptide Agonists,V.J. Hruby, R.S. Agnes, C. Cai.Design of Nonpeptides from Peptide Ligands for Peptide Receptors,V.J. Hruby, W. Qiu, T. Okayama, and V.A. Soloshonok.Strategies for Mapping the Binding Site of the Serotonin 5-HT2A Receptor,B.J. Ebersole and S.C. Sealfon.C. Structural Characterization of Receptor Proteins:Use of the Substituted Cysteine Accessibility Method to Study the Structure and Function of G Protein-Coupled Receptors,J.A. Javitch, L. Shi, and G. Liapakis.Mass Spectrometric Analysis of G Protein-Coupled Receptors,D.R. Knapp, R.K. Crouch, L.E. Ball, A.K. Gelasco, and Z. Ablonczy.Probing the Higher Order Structure of G protein-Coupled Receptors Using Tethered Cleavage Methods,A.K. Gelasco, R.K. Crouch, and D.R. Knapp.Use of Fluorescence Spectroscopy to Study Conformational Changes in the b2-Adrenoceptor,B.K. Kobilka and U. Gether.Crystallization of Membrane Protein in Cubo,P. Nollert, J. Navarro, and E.M. Landau.N-Linked Carbohydrates on G Protein-Coupled Receptors: Mapping Sites of Attachment and Determining Functional Roles,D.P. Davis and D.L. Segaloff.Magic Angle Spinning Nuclear Magnetic Resonance of Isotropically Labeled Rhodopsin,M. Eilers, W. Ying, P.J. Reeves, H.G. Khorana, and S.O. Smith.Use of Nuclear Magnetic Resonance to Study the Three-Dimensional Structure of Rhodopsin,P.L. Yeagle and A.D. Albert.D. Design and Use of Engineered Receptor Proteins:Tools for Dissecting Signaling Pathways in Vivo: Receptors Activated Solely by Synthetic Ligands,K. Scearce-Levie, P. Coward, C.H. Redfern, and B.R. Conklin.Analysis of Structure-Function from Expression of G Protein-Coupled Receptor Fragments,S.S. Karnik.Construction and Analysis of Function of G Protein-Coupled Receptor-G Protein Fusion Proteins,G. Milligan.Synthetic Gene Technology: Applications to Ancestral Gene Reconstruction and Structure-Function Studies of Receptors,B.S.W. Chang, M.A. Kazmi, and T.P. Kakmar.Considerations in the Design and Use of Chimeric G Protein-Coupled Receptor,J. Wess.E. Molecular Modeling Studies of Receptor Structure and Function:Strategies for Modeling the Interactions of Transmembrane Helices of G Protein-Coupled Receptors by Geometric Complementarity Using the GRAMM Computer Algorithm,I.A. Vakser and S. Jiang.Three-Dimensional Representation of G Protein-Coupled Receptor Structures and Mechanisms,I. Visiers, J.A. Ballesteros, and H. Weinstein.F. Analysis of Receptor Protein Coupling:Reconstitution of G Protein-Coupled Receptors with Recombinant G Protein a and bg Subunits,W.E. McIntire, C.S. Myung, G. MacCleery, Q. Wang, and J.C. Garrison.Cell-Free Membrane Desensitization Assay for G Protein-Coupled Receptors,M. Hunzicker-Dunn and L. Birnbaumer.Methods to Determine the Constitutive Activity of Histamine H2 Receptors,R. Leurs, M. Hoffman, A.E. Alewijnse, M.J. Smit, and H. Timmerman.Expression of G Protein-Coupled Receptors and G Proteins in Sf9 Cells: Analysis of Coupling by Radioligand Binding,R.T. Windh and D.R. Manning.G Protein-Coupled Receptors and Proliferative Signaling,M.J. Smit, R.A. Bakker, and E.S. Burstein.G. Characterization of Receptor Heterogeneity:Genetic Analysis of G Protein-Coupled Receptor Genes,D.P. Healy.Identification of Adrenergic Receptor Polymorphisms,K.M. Small, D.A. Rathz, and S.B. Liggett.Strategies and Requirements for the Detection of RNA Editing in G Protein Coupled-Receptor RNA,C.M. Niswender.H. The Study of Receptor Trafficking:Fluorescence Microscopy Techniques for the Study of G Protein-Coupled Receptor Trafficking,L. Kallal and J.L. Benovic.Measurement of Receptor Desensitization and Internalization in Interact Cells,R.B. Clark and B.J. Knoll.Morphological and Biochemical Strategies for Monitoring Trafficking of Epitope-Tagged G Protein-Coupled Receptors in Agonist-Naïve and Agonist-Occupied States,M. Wozniak, C. Saunders, N. Schramm, J.R. Keefer, and L.E. Limbird.Section II. Regulators of GPCR Function.A. G Protein-Coupled Receptor Kinases (GRKs):Characterization of G Protein-Coupled Receptor Kinases,A.N. Pronin, R.P. Loudon, and J.L. Benovic.Regulation of G Protein-Coupled Receptor Kinase 2,T. Haga, K. Haga, K. Kameyama, H. Tsuga, and N. Yoshida.Rhodopsin and Its Kinase,I. Sokal, A. Pulvermuller, J. Buczylko, K.P. Hofmann, and K. Palczewski.B. Arrestins and Novel Proteins:Characterization of Arrestin Expression and Function,S.J. Mundell, M.J. Orsini, and J.L. Benovic.Identification of Novel G Protein-Coupled Receptor-Interacting Proteins,R.T. Premont and R.A. Hall.Author Index.Subject Index.