Synthesis and application of a novel GTP phosphonamide photoaffinity reagent: Study of exchangeable GTP-binding domain on tubulin

Thesis/Dissertation · Sun Jan 01 04:00:00 EST 1989

OSTI ID:5605031

Chavan, A J

The regulatory role played by nucleotides in various biochemical processes has been the topic of study for several years. This has led to the identification of several regulatory proteins which require guanosine triphosphate (GTP), a nucleotide, for their action. The G-protein family utilizes GTP and displays many common features in the mechanism of their action. Chapter 1 is an overview of several GTP-binding proteins including G-proteins. Tubulin, a structural protein also requires GTP for its assembly/disassembly process. The nature of interaction of GTP with tubulin, particularly at the exchangeable GTP-binding site has been under investigation for several years. Chapter 2 presents an overview of the structure of tubulin, and the studies leading up to the present understanding of the GTP-tubulin interaction. To study the exchangeable GTP-binding domain on tubulin which would complement the other techniques, and reagents used, the author synthesized a novel GTP phosphonamide photoaffinity reagent. Chapter 3 describes the synthesis of {sup 125}I-APTG (36). He utilized {sup 125}I-APTG to establish that it could act as a mimic to GTP. Using the combination of anion-exclusion chromatography, gel-filtration and reverse-phase HPLC techniques he isolated residues 65-79 on the {beta}-chain as the major peptide in the GTP-binding domain. He also isolate residues 65-79 and 353-370 on the {alpha}-chain as minor peptides supporting the hypothesis that the phosphate region is in close proximity of the {alpha}-subunit.

• Research Organization: Kentucky Univ., Lexington, KY (United States)
• OSTI ID: 5605031
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BIOLOGICAL FUNCTIONS
CHEMICAL PROPERTIES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE 125
IODINE ISOTOPES
ION EXCHANGE CHROMATOGRAPHY
ISOTOPE APPLICATIONS
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
MEMBRANE PROTEINS
MOLECULAR STRUCTURE
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
REAGENTS
RECEPTORS
SEPARATION PROCESSES
TRACER TECHNIQUES