Abstract:
:Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy has emerged as an efficient tool to elucidate the structure and the conformational dynamics of proteins under conditions close to the native state. This review article summarizes the basics as well as the recent progress in SDSL and EPR methods, especially for investigations on protein structure, protein function, and interaction of proteins with other proteins or nucleic acids. Labeling techniques as well as EPR methods are introduced and exemplified with applications to systems that have been studied in the author's laboratory in the past 15 years, headmost the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis. Further examples underline the application of SDSL EPR spectroscopy to answer specific questions about the system under investigation, such as the nature and influence of interactions of proteins with other proteins or nucleic acids. Finally, it is discussed how SDSL EPR can be combined with other biophysical techniques to combine the strengths of the different methodologies.
journal_name
Biol Chemjournal_title
Biological chemistryauthors
Klare JPdoi
10.1515/hsz-2013-0155subject
Has Abstractpub_date
2013-10-01 00:00:00pages
1281-300issue
10eissn
1431-6730issn
1437-4315pii
/j/bchm.just-accepted/hsz-2013-0155/hsz-2013-0155.journal_volume
394pub_type
杂志文章,评审abstract::Adenoviral DNA was examined within capsids by dimethyl sulfate footprinting. Protein-DNA interactions were visualized through ligation-mediated PCR (LM-PCR). Signals for protein binding were found adjacent to both inverted terminal repeats (ITR). There were no indications of close protein binding at several other loci...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2001.169
更新日期:2001-09-01 00:00:00
abstract::The enzymatic properties of glycosylphosphatidylinositol-specific phospholipase D (EC 3.1.4.50) were characterized using a 6,000-fold purified enzyme. This was obtained in 100 microg amounts from human serum with a recovery of 35%. Pure alkaline phosphatase containing one anchor moiety per molecule was used as substra...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2000.062
更新日期:2000-05-01 00:00:00
abstract::The purpose of this work was to study the regulative role of the glutamate receptor found earlier in the brain mitochondria. In the present work a glutamate-dependent signaling system with similar features was detected in mitochondria of the heart. The glutamate-dependent signaling system in the heart mitochondria was...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2015-0289
更新日期:2016-05-01 00:00:00
abstract::We present a rapid and simple method for the sexing of human and other primate DNA. Homologous introns from the X- and Y-linked zinc finger protein genes (ZFX and ZFY) are PCR amplified with one primer pair. An Alu element insertion in the ZFX intron results in a size difference in the amplicons which is resolved by a...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:
更新日期:1998-10-01 00:00:00
abstract::Cytokinins are essential plant hormones that regulate numerous physiological processes. Recently, a protein was identified in mung bean (Vigna radiata) and characterized as a cytokinin-specific binding protein (VrCSBP). Fluorescence correlation spectroscopy was used to investigate the interaction between VrCSBP and it...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2010.005
更新日期:2010-01-01 00:00:00
abstract::The sphingolipid sphingosine-1-phosphate (S1P) has various functions in immune cell biology, regulating survival, proliferation, and, most prominently, migration. S1P couples to five G protein-coupled receptors (S1PR1-5) to transduce its effects on immune cell function. Expression of S1PR4 is restricted to immune cell...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2014-0271
更新日期:2015-06-01 00:00:00
abstract::Host T cell activation, a key step in obtaining adaptive immunity against pathogens, is initiated by the binding of the T cell receptor to a foreign antigenic peptide presented by the major histocompatibility complex on the surface of an antigen-presenting cell and, consequently, formation of an immunological synapse....
journal_title:Biological chemistry
pub_type: 杂志文章,评审
doi:10.1515/hsz-2014-0282
更新日期:2015-06-01 00:00:00
abstract::We have previously shown that short double-stranded RNAs of specific sequence induce phosphorylation in the activation loop of splicing variant D of the transforming growth factor beta-activated protein kinase 1 (TAK1-D). Here, we further characterize this novel function of TAK1-D and the mechanisms of this dsRNA-trig...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2009.046
更新日期:2009-05-01 00:00:00
abstract::The average amount of cholesterol in the whole animal equals approximately 2100 mg/kg body weight, and 15% and 23% of this sterol in the mouse and human, respectively, is found in the central nervous system. There is no detectable uptake across the blood-brain barrier of cholesterol carried in lipoproteins in the plas...
journal_title:Biological chemistry
pub_type: 杂志文章,评审
doi:10.1515/BC.2009.035
更新日期:2009-04-01 00:00:00
abstract::The immunoproteasome subunit low molecular weight protein 2 (LMP2) codon 60 polymorphism has been associated with autoimmune diseases. It has also been demonstrated to influence susceptibility to TNF-alpha-induced apoptosis in blood cells and proteasome activity in aged human brain. In the present study, an in silico ...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2006.056
更新日期:2006-04-01 00:00:00
abstract::The conversion of the cellular isoform of the prion protein (PrP(C)) into the pathologic isoform (PrP(Sc)) is the key event in prion diseases. To study the conversion process, an in vitro system based on varying the concentration of low amounts of sodium dodecyl sulfate (SDS) has been employed. In the present study, t...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2011.048
更新日期:2011-05-01 00:00:00
abstract::Hypoxia-inducible transcription factor (HIF), an αβ dimer, is the key inducer of hypoxia-responsive genes that operate both during normal development and pathological processes in association with decreased oxygen availability. The products of HIF target genes function in, e.g., hematopoiesis, angiogenesis, iron trans...
journal_title:Biological chemistry
pub_type: 杂志文章,评审
doi:10.1515/hsz-2012-0328
更新日期:2013-04-01 00:00:00
abstract::Dendritic cells (DCs) are an important link between innate and adaptive immunity. DCs get activated in inflamed tissues where oxygen tension is usually low, which requires the transcription factor hypoxia inducible factor (HIF)-1 for cellular adaptation. To investigate whether the HIF-1 transcriptional complex plays a...
journal_title:Biological chemistry
pub_type: 杂志文章,评审
doi:10.1515/hsz-2012-0320
更新日期:2013-04-01 00:00:00
abstract::Abstract The R- and K-gingipain proteases of Porphyromonas gingivalis are involved in proteolysis of haemoglobin from which the defensive dimeric haem pigment is formed. Whilst oxyhaemoglobin is refractory towards K-gingipain, methaemoglobin is rapidly degraded. Ligation of methaemoglobin with N3-, which effectively b...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2008.140
更新日期:2008-09-01 00:00:00
abstract::The influence of the cellular milieu, a complex and crowded solvent, is often neglected when biomolecular structure and function are studied in vitro. To mimic the cellular environment, crowding effects are commonly induced in vitro using artificial crowding agents like Ficoll or dextran. However, it is unclear if suc...
journal_title:Biological chemistry
pub_type: 杂志文章,评审
doi:10.1515/hsz-2015-0161
更新日期:2016-01-01 00:00:00
abstract::Animal cells release traces of material onto glass or silicon surfaces during adhesion and migration. This little studied phenomenon is a widespread and normal concomitant of cell migration. The paper introduces the study of such material. The traces can be visualised by different microscopic techniques (e.g. TIRF, IR...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/bchm.1998.379.8-9.1161
更新日期:1998-08-01 00:00:00
abstract::Arginine deiminase (ADI) is a potential antitumor agent for the arginine deprivation treatment of L-arginine auxotrophic tumors. The optimum pH of ADI varies significantly, yet little is known about the origin of this variety. Here, Pseudomonas aeruginosa ADI (PaADI), an enzyme that functions only at acidic pH, was ut...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2012-0183
更新日期:2012-09-01 00:00:00
abstract::MHC encoded DM heterodimers and classical MHC class II complexes meet in an endosomal/lysosomal compartment where DM heterodimers support peptide loading of MHC class II. Studies on peptide loading of rat class II and on peptide persistence in cells of the dendritic lineage prompted us to establish full length cDNA cl...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/bchm.1997.378.9.1005
更新日期:1997-09-01 00:00:00
abstract::We have previously shown that compatible organic osmolytes, such as betaine, myo-inositol and taurine, are part of the stress response of normal human keratinocytes (NHKs) to ultraviolet B (UVB) radiation. In this regard, we tested human HaCaT keratinocytes as a surrogate cell line for NHK. HaCaT cells osmo-dependentl...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2007.140
更新日期:2007-12-01 00:00:00
abstract::Thyroid hormones stimulate gap junctional communication in rat liver WB-F344 epithelial cells, elevating connexin43 mRNA and protein levels. In the present work we analysed connexin43 expression in liver and heart samples from thyroid hormone-treated Wistar rats. Connexin43 mRNA was elevated 2.1-fold in rat liver samp...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2000.120
更新日期:2000-09-01 00:00:00
abstract::FK506-binding proteins (FKBPs) belong to a distinct class of immunophilins that interact with immunosuppressants. They use their peptidyl-prolyl isomerase (PPIase) activity to catalyze the cis-trans conversion of prolyl bonds in proteins during protein-folding events. FKBPs also act as a unique group of chaperones. Th...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2017-0251
更新日期:2018-04-25 00:00:00
abstract::Rasal is a modular multi-domain protein of the GTPase-activating protein 1 (GAP1) family; its four known members, GAP1m, Rasal, GAP1IP4BP and Capri, have a Ras GTPase-activating domain (RasGAP). This domain supports the intrinsically slow GTPase activity of Ras by actively participating in the catalytic reaction. In t...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2017-0159
更新日期:2017-12-20 00:00:00
abstract::The physiological function of the GPI-anchored ectoenzyme aminopeptidase P (APP) is still elusive. Most researchers suppose that this enzyme inactivates biologically active peptides like bradykinin, neuropeptide tyrosine (NPY) and others (Vanhoof et al., 1995). We demonstrate by immunohistology with a specific antibod...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/bchm.1998.379.6.705
更新日期:1998-06-01 00:00:00
abstract::The glutamyl endopeptidase family of enzymes from staphylococci has been shown to be important virulence determinants of pathogenic family members, such as Staphylococcus aureus. Previous studies have identified the N-terminus and residues from positions 185-195 as potentially important regions that determine the acti...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2010.116
更新日期:2010-10-01 00:00:00
abstract::Metachromatic leukodystrophy is a lysosomal disorder caused by the deficiency of arylsulfatase A (ASA). This leads to the storage of the sphingolipid 3-O-sulfogalactosylceramide (sulfatide) in various cell types, such as renal tubular cells. Examination of mannose 6-phosphate receptor (MPR300)-dependent endocytosis re...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2009.009
更新日期:2009-01-01 00:00:00
abstract::N-acyl homoserine lactones (AHLs) are quorum sensing (QS) signal molecules used by most Gram-negative pathogenic bacteria. In this article the lactonase activity of the preparations based on hexahistidine-tagged organophosphorus hydrolase (His6-OPH) towards AHLs was studied. Initially, three of the most interesting β-...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2018-0162
更新日期:2018-07-26 00:00:00
abstract::Interleukin-1 signaling is initiated by recruitment of adapter proteins and kinases to the type I interleukin-1 receptor (IL-1RI). It is modulated by accompanying redox processes at various levels, such as (auto-) phosphorylation of the IL-1RI-associated kinase IRAK, the phosphorylation of IkappaB and translocation an...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2003.068
更新日期:2003-04-01 00:00:00
abstract::Major vault protein (MVP) is the predominant member of a large cytosolic ribonucleoprotein particle, termed vault. We have previously shown that MVP derived from electric ray electric organ becomes phosphorylated by protein kinase C in vitro and by tyrosine kinase in vivo. Here we show that MVP from two mammalian cell...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/BC.2001.180
更新日期:2001-10-01 00:00:00
abstract::During the last years, the use of therapeutic glycoproteins has increased strikingly. Glycosylation of recombinant glycoproteins is of major importance in biotechnology, as the glycan composition of recombinant glycoproteins impacts their pharmacological properties. The terminal position of N-linked complex glycans in...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2012-0138
更新日期:2012-08-01 00:00:00
abstract::As an important protein, UL31 has been demonstrated to play multiple roles in herpes simplex virus 1 (HSV-1) replication. Previous studies showed that UL31 predominantly locates in the nucleus in chemical fixed cells and live cells, however, the determining mechanisms for its nuclear translocation is not clear. In the...
journal_title:Biological chemistry
pub_type: 杂志文章
doi:10.1515/hsz-2015-0299
更新日期:2016-06-01 00:00:00