Conformational state of a 25-mer peptide from the cyclophilin-binding loop of the HIV type 1 capsid protein

Reimer, Ulf; Drewello, Mario; Jakob, Mario; Fischer, Gunter; Schutkowski, Mike

doi:10.1042/bj3260181

Cited by 15 publications

(15 citation statements)

References 31 publications

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“…Isomerization around L-P bonds corresponding to the L19-P20 and L644-P645 positions in the full length TRPC1 was slow on the NMR timescale (< 0.1 s −1 ), as shown by distinct amide resonance signals corresponding to cis and trans conformations in approximately 1:10 ratio consistent with thermodynamically expected distribution of the two conformers for an unstructured peptide (Reimer et al, 1997) (Figure 1D). To assess the effect of FKBPs on TRPC1, we performed 2-dimentional 1 H- 15 N heteronuclear exchange experiments (Farrow et al, 1994) on 15 N-labelled TRPC1(14–30) and TRPC1(635–656) peptides in the presence of catalytic amounts of recombinant FKBP12 or FKBP52.…”

Section: Resultsmentioning

confidence: 64%

Peptidyl-Prolyl Isomerase FKBP52 Controls Chemotropic Guidance of Neuronal Growth Cones via Regulation of TRPC1 Channel Opening

Shim

Yuan

Kim

et al. 2009

Neuron

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Summary Immunophilins, including FK506-binding proteins (FKBPs), are protein chaperones with peptidyl-prolyl isomerase (PPIase) activity. Initially identified as pharmacological receptors for immunosuppressants to regulate immune responses via isomerase independent mechanisms, FKBPs are most highly expressed in the nervous system where their physiological function as isomerases remains unknown. We demonstrate that FKBP12 and FKBP52 catalyze cis/trans isomerization of regions of TRPC1 implicated in controlling channel opening. FKBP52 mediates stimulus-dependent TRPC1 gating through isomerization, which is required for chemotropic turning of neuronal growth cones to netrin-1 and myelin-associated glycoprotein and for netrin-1/DCC-dependent midline axon guidance of commissural interneurons in the developing spinal cord. By contrast, FKBP12 mediates spontaneous opening of TRPC1 through isomerization and is not required for growth cone responses to netrin-1. Our study demonstrates a novel physiological function of proline isomerases in chemotropic nerve guidance through TRPC1 gating and may have significant implication in clinical applications of immunophilin-related therapeutic drugs.

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Section: Resultsmentioning

confidence: 64%

Peptidyl-Prolyl Isomerase FKBP52 Controls Chemotropic Guidance of Neuronal Growth Cones via Regulation of TRPC1 Channel Opening

Shim

Yuan

Kim

et al. 2009

Neuron

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“…Originally, prolyl cis/trans isomerase activity of CypA with relatively short model peptides, containing one Pro residue, was revealed by line shape analysis of 1 D spectra [ 31 - 33 ]. Exchange spectroscopy including 2D 1 H- 1 H NOESY and ROESY NMR experiments have been applied to determine prolyl cis/trans isomerase interactions of CypA with unlabelled model peptides [ 23 , 34 , 35 ]. Selective interaction of Pro-90 of HIV-1 Capsid was revealed by 3D 15 N-edited NOESY-HSQC of 15 N-labelled N-terminal (1-146) and full-length HIV-1 Capsid, in addition to a fragment of the HIV-1 Gag polyprotein containing the full Matrix protein and the N-terminal domain of Capsid [ 12 , 13 ].…”

Section: Resultsmentioning

confidence: 99%

The intriguing Cyclophilin A-HIV-1 Vpr interaction: prolyl cis/trans isomerisation catalysis and specific binding

et al. 2010

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BackgroundCyclophilin A (CypA) represents a potential target for antiretroviral therapy since inhibition of CypA suppresses human immunodeficiency virus type 1 (HIV-1) replication, although the mechanism through which CypA modulates HIV-1 infectivity still remains unclear. The interaction of HIV-1 viral protein R (Vpr) with the human peptidyl prolyl isomerase CypA is known to occur in vitro and in vivo. However, the nature of the interaction of CypA with Pro-35 of N-terminal Vpr has remained undefined.ResultsCharacterization of the interactions of human CypA with N-terminal peptides of HIV-1 Vpr has been achieved using a combination of nuclear magnetic resonace (NMR) exchange spectroscopy and surface plasmon resonance spectroscopy (SPR). NMR data at atomic resolution indicate prolyl cis/trans isomerisation of the highly conserved proline residues Pro-5, -10, -14 and -35 of Vpr are catalyzed by human CypA and require only very low concentrations of the isomerase relative to that of the peptide substrates. Of the N-terminal peptides of Vpr only those containing Pro-35 bind to CypA in a biosensor assay. SPR studies of specific N-terminal peptides with decreasing numbers of residues revealed that a seven-residue motif centred at Pro-35 consisting of RHFPRIW, which under membrane-like solution conditions comprises the loop region connecting helix 1 and 2 of Vpr and the two terminal residues of helix 1, is sufficient to maintain strong specific binding.ConclusionsOnly N-terminal peptides of Vpr containing Pro-35, which appears to be vital for manifold functions of Vpr, bind to CypA in a biosensor assay. This indicates that Pro-35 is essential for a specific CypA-Vpr binding interaction, in contrast to the general prolyl cis/trans isomerisation observed for all proline residues of Vpr, which only involve transient enzyme-substrate interactions. Previously suggested models depicting CypA as a chaperone that plays a role in HIV-1 virulence are now supported by our data. In detail the SPR data of this interaction were compatible with a two-state binding interaction model that involves a conformational change during binding. This is in accord with the structural changes observed by NMR suggesting CypA catalyzes the prolyl cis/trans interconversion during binding to the RHFP35RIW motif of N-terminal Vpr.

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“…Thus, non‐specific binding cannot be identified as the cause of insensitivity to catalytic interconversions. Since ‐Gly‐Pro‐ peptides can be effectively interconverted by Cyp18 [42]the Ala side chain methyl preceding proline is not absolutely required to assist the activation of the catalytic machinery of the PPIase. Taken together these results point out that a ground state P 1 diastereomeric complex may be resistant to undergo considerable spatial rearrangements toward of the transition state due to steric hindrance of the wrongly positioned Ala side chain.…”

Section: Discussionmentioning

confidence: 99%

Mapping the stereospecificity of peptidyl prolyl cis/trans isomerases

et al. 1998

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The stereospecificity of peptidyl prolyl cis/trans isomerases (PPIases) was studied using tetrapeptide substrate analogs in which one amino acid residue was replaced by the cognate D-amino acid in various positions of the peptide chain. Reversed stereocenters around proline markedly increased the rate of the spontaneous trans to cis isomerization of the prolyl bond whereas cis to trans isomerizations were less sensitive. PPIases like human cyclophilin18, human FKBP12, Escherichia coli parvulin10 and the PPIase domain of E. coli trigger factor exhibited stereoselectivity demanding at the P I to P P P position of the substrate chain. The discriminating factor for stereoselectivity was the lack of formation of the Michaelis complexes of the diastereomeric substrates. However, D-alanine at the P I position preserved considerable affinity to the active site, and largely prevented activation of the catalytic machinery for all PPIases investigated.z 1998 Federation of European Biochemical Societies.

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Conformational state of a 25-mer peptide from the cyclophilin-binding loop of the HIV type 1 capsid protein

Cited by 15 publications

References 31 publications

Peptidyl-Prolyl Isomerase FKBP52 Controls Chemotropic Guidance of Neuronal Growth Cones via Regulation of TRPC1 Channel Opening

Peptidyl-Prolyl Isomerase FKBP52 Controls Chemotropic Guidance of Neuronal Growth Cones via Regulation of TRPC1 Channel Opening

The intriguing Cyclophilin A-HIV-1 Vpr interaction: prolyl cis/trans isomerisation catalysis and specific binding

Mapping the stereospecificity of peptidyl prolyl cis/trans isomerases

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