Identification of functionally important acidic residues in transducin by group-specific labeling

2Citations
Citations of this article
6Readers
Mendeley users who have this article in their library.

Abstract

Transducin (T), a GTP-binding protein involved in phototransduction of rod photoreceptor cells, is a heterotrimer arranged as two units, the α-subunit (T α ,) and the βγ-complex (T βγ ). The role of the carboxyl groups in T was evaluated by labeling with N, N′-dicyclohexylcarbodiimide (DCCD) and 1-ethyl 3-(3-dimethylaminopropyl) carbodiimide (EDC). Only a minor effect on the binding of β, γ-imido guanosine 5′-triphosphate (GMPpNp to T was observed in the presence of the hydrophobic carbodiimide, DCCD. Similarly, the GMPpNp binding activity of the reconstituted holoenzyme was not significantly affected when T α was combined with DCCD-treated T βγ . However, the binding of guanine nucleotides to the reconstituted T was ∼50% inhibited when DCCD-labeled Tα was incubated with T βγ . In contrast, treatment of T with the hydrophilic carbodiimide, EDC, completely impaired its GMPpNp-binding ability. EDC-modified T was incapable of interacting with illuminated rhodopsin, as determined by sedimentation experiments. However, rhodopsin only partially protected against the inactivation of T. Additionally, analyses of trypsin digestion patterns showed that fluoroaluminate was not capable of activating the EDC-labeled T sample. The function of the reconstituted holoenzyme was also disrupted when EDC-modified T α was combined with T βγ , and when EDC-treated T βγ was incubated with T α .

Cite

CITATION STYLE

APA

Kosoy, A., Möller, C., Perdomo, D., & Bubis, J. (2003). Identification of functionally important acidic residues in transducin by group-specific labeling. Biological Research, 36(3–4), 389–404. https://doi.org/10.4067/S0716-97602003000300010

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free