Abstract
Nuclear magnetic resonance spectroscopic structure determination of proteins has been under rapid development during the last decade. The size limitation impeding
structural studies of biological macromolecules in solution has increased from 10 kDa to 30 kDa thanks to exploitation of
15N/13C enrichment. Perdeuteration of non-exchangeable protons has pushed this limit even further, allowing backbone
resonance assignment of 40 to 50 kDa proteins. Most recently, transverse relaxation optimized spectroscopy (TROSY) has been demonstrated to lengthen
15N and 1HN spin transverse relaxation times significantly,
especially in large perdeuterated proteins, thus extending the size limit beyond 100 kDa systems. However, determination of structurally important nuclear Overhauser
enhancements (NOE)
suffers from perdeuteration, due to the lower density of proton spins available, eventually leading to imprecise protein structures. Very recently, residual dipolar
couplings have been used to supplement NOE information, enabling accurate molecular structures to also be obtained with perdeuterated proteins. This thesis focuses on the
measurement of the structurally important 3J-coupling between 1HN and
1Hα spins, and determination of residual dipolar couplings by utilizing the novel spin-state-selective
subspectral editing together with the TROSY methodology. This approach allows precise measurement of a large number of dipolar couplings in larger protonated or
perdeuterated proteins.
| Identifer | oai:union.ndltd.org:oulo.fi/oai:oulu.fi:isbn951-42-5822-3 |
| Date | 03 November 2000 |
| Creators | Permi, P. (Perttu) |
| Publisher | University of Oulu |
| Source Sets | University of Oulu |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/doctoralThesis, info:eu-repo/semantics/publishedVersion |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess, © University of Oulu, 2000 |
| Relation | info:eu-repo/semantics/altIdentifier/pissn/0355-3191, info:eu-repo/semantics/altIdentifier/eissn/1796-220X |
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