Complete 1H, 13C, and 15N assignments and secondary structure of the GTPase activating domain of Gs

Biochemistry. 1995 Jan 10;34(1):155-62. doi: 10.1021/bi00001a019.

Abstract

Complete 1H, 13C, and 15N assignments for backbone and side-chain atoms of the 145 residue GTPase activating domain of Gs are presented. The combination of gradient-enhanced versions of the HNCACB and CBCA(CO)NNH pulse sequences provided enough information to obtain sequential backbone assignments for residues 2-145 of the polypeptide, as well as assignments of asparagine and glutamine side-chain amides. HBHA(CO)NNH, HCCH-TOCSY, and 13C/15N NOESY-HSQC experiments yielded side-chain 1H and 13C assignments. Chemical shift data and 15N NOESY-HSQC experiments provided information on the secondary structure of the domain, which is similar to that observed in the cognate domain in transducin, a related G protein. The functionally essential C-terminal 15 residues are disordered in solution. These assignments provide a basis for determining the solution structure of the domain.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Carbon Isotopes
  • GTP-Binding Proteins / chemistry*
  • GTPase-Activating Proteins
  • Hydrogen
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Nitrogen Isotopes
  • Protein Structure, Secondary*
  • Proteins / chemistry*

Substances

  • Carbon Isotopes
  • GTPase-Activating Proteins
  • Nitrogen Isotopes
  • Proteins
  • Hydrogen
  • GTP-Binding Proteins