Synthetic peptides as probes for conformational preferences of domains of membrane receptors.
Peptide models have been widely used to investigate conformational aspects of domains of proteins since the early 1950s. A pioneer in this field was Dr. Murray Goodman, who applied a battery of methodologies to study the onset of structure in homooligopeptides. This article reviews some of Dr. Goodman's contributions, and reports recent studies using linear and constrained peptides corresponding to the first extracellular loop and linear peptides corresponding to the sixth transmembrane domain of a G-protein coupled receptor from the yeast Saccharomyces cerevisiae. Peptides containing 30-40 residues were synthesized using solid-phase methods and purified to near homogeneity by reversed phase high performance liquid chromatography. CD and NMR analyses indicated that the first extracellular loop peptides were mostly flexible in water, and assumed some helical structure near the N-terminus in trifluoroethanol and in the presence of micelles. Comparison of oligolysines with native loop residues revealed that three lysines at each terminus of a peptide corresponding to the sixth transmembrane domain of the alpha-factor receptor resulted in better aqueous solubility and greater helicity than the native loop residues.
Naider
Fred
F
Department of Chemistry, College of Staten Island, and Institute for Macromolecular Assemblies, City University of New York, Staten Island, NY 10314, USA. Naider@mail.csi.cuny.edu
Khare
Sanjay
S
Arshava
Boris
B
Severino
Beatrice
B
Russo
Joe
J
Becker
Jeffrey M
JM
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D049673:History, 20th Century; D008958:Models, Molecular; D015335:Molecular Probes / Q000266:history; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000138:chemical synthesis / Q000266:history; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000266:history; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide / Q000737:chemistry / Q000266:history; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000266:history; D014157:Transcription Factors / Q000737:chemistry / Q000266:history
D015335:Molecular Probes; D010455:Peptides; D011956:Receptors, Cell Surface; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors