Mapping the stereospecificity of peptidyl prolyl <i>cis/trans</i> isomerases

Schiene, Cordelia; Reimer, Ulf; Schutkowski, Mike; Fischer, Gunter

doi:10.1016/s0014-5793(98)00871-0

Cited by 31 publications

(40 citation statements)

References 46 publications

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“…The oligopeptide conformation is known to tolerate the prolyl peptide bond surrogate Xaa-[CS-N]-Pro, but on the contrary, the catalysis by Cyp18-like PPIases is abolished (27). Notably, a conservation of ground state binding affinity of oligopeptides with a changed stereochemistry on the C-R atom at the P1 position to various PPIases has been reported (28). However, the enzymatic activities of these PPIases are highly stereospecific.…”

Section: Resultsmentioning

confidence: 99%

Substrate-Based Design of Reversible Pin1 Inhibitors

et al. 2002

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Human Pin1, a peptidyl-prolyl cis/trans isomerase with high specificity to -Ser/Thr(PO(3)H(2))-Pro- motifs, is required for cell cycle progression. In an effort to design reversible Pin1 inhibitors by using a substrate structure based approach, a panel of peptides were applied to systematically analyze the minimal structural requirements for Pin1 substrate recognition. Pin1 catalysis (k(cat)/K(m) < 5 mM(-1) s(-1)) for Ala-Pro, Ser-Pro, and Ser(PO(3)H(2))-Pro was detected using direct UV-visible spectrophotometric detection of prolyl isomerization, while weak competitive inhibition of Pin1 by these dipeptides was observed (K(i) > 1 mM). Substrates with chain lengths extending from either the P2 to P1' or the P1 to P2' subsite gave k(cat)/K(m) values of 100 mM(-1) s(-1) for Ala-Ser(PO(3)H(2))-Pro and 38 mM(-1) s(-1) for Ser(PO(3)H(2))-Pro-Arg. For both Pin1 and its yeast homologue Ess1, the optimal subsite recognition elements comprise five amino acid residues with the essential Ser(PO(3)H(2)) in the middle position. The resulting substrate Ac-Ala-Ala-Ser(PO(3)H(2))-Pro-Arg-NH-4-nitroanilide possesses a very low cis/trans interconversion barrier in the presence of either Pin1 or Ess1, with k(cat)/K(m) = 9300 mM(-1) s(-1) and 12000 mM(-1) s(-1), respectively. The D-Ser(PO(3)H(2)) residue preceding proline could serve as a substrate-deactivating determinant without compromising ground state affinity. Similarly, substitution of the amide bond preceding proline with a thioxo amide bond produces a potent inhibitor. Pin1 is reversibly inhibited by such substrate analogue inhibitors with IC(50) values in the low micromolar range. The D-amino acid containing inhibitor also exhibits remarkable stability against phosphatase activity in cell lysate.

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Section: Resultsmentioning

confidence: 99%

Substrate-Based Design of Reversible Pin1 Inhibitors

et al. 2002

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“…Various methodologies for measuring PPIase activity have been described by many authors [2,91,[94][95][96][97][98] who used linear tetrapeptides containing a proline residue, for example Suc-[Yaa] n -Xaa-Pro-Phe-pNA. The X-Pro bond of the tetrapeptides dissolved in aqueous solution is in equilibrium between the cis and trans forms.…”

Section: Ppiase Activity and Its Attributesmentioning

confidence: 99%

Peptidylprolyl Cis / Trans Isomerases (Immunophilins): Biological Diversity - Targets - Functions

Gałat

2003

CTMC

344

302

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Information recovered from genome sequencing projects, multiple sequence alignments, structural analyses of PPIase and published records were used in deciphering the biological diversity, functions and targets of four groups of proteins encoded by dissimilar sets of sequences whose spatial representations exhibit peptidylprolyl cis/trans isomerase activity (PPIase). In the human genome there are encoded fifteen proteins whose segments have significant homology with the sequence of 12 kDa protein which is the target of the potent immunosuppressive macrolides FK506 or rapamycin. The 12 kDa archetype of the FK506-binding protein (FKBP), known as FKBP-12a, is an abundant intracellular protein whereas other FKBPs possessing from one to four FK506-like binding domains (FKBDs) have nominal masses varying from 13 to 135 kDa. The human genome contains at least sixteen genes encoding proteins comprising one cyclosporin-A (CsA) binding domain (CLD) called cyclophilins whose nominal masses vary from 17 to 324 kDa and multiple coding segments for small cyclophilins (17-19 kDa) whose transcription levels and functions remain unknown. The third group of PPIases encoded in the genome comprises two proteins (hPin1 and hParv14) where hPin1 is an important PPIase for cell cycle. The A. thaliana, C. elegans, D. melanogaster and S. cerevisiae genomes encode a less diverse spectrum of PPIases whereas the prokaryotic genomes contain from none to three cyclophilins, from none to four genes encoding FKBPs, one distant homologue of the Pin1 protein named parvulin and the fourth group of PPIases known as trigger factors. PPIases are discretely distributed to different cellular compartments and interact with a number of targets that control a range of cellular processes. Analyses of the sequence alignments of the two groups of PPIases, namely cyclophilins and FKBPs from diverse phyla, show that in each group their sequences diverge but the amino acid residues which form the PPIase activity site and macrolide binding cavity remain well conserved in the majority of them which suggests that the spatial structures and functions of each group of PPIases remain conserved.

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“…Resolution of several structures of peptide:cyclophilin complexes [10–13] as well as kinetic studies have shown that the proline residue with the ( L ) configuration is essential for the PPIase activity [13,14] although it may be replaced with proline isosteres [15]. The influence of other residues of the substrate on the interaction is not so critical.…”

Section: Introductionmentioning

confidence: 99%

Interaction of human cyclophilin hCyp‐18 with short peptides suggests the existence of two functionally independent subsites

et al. 2001

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The binding of peptides, derived from the model substrate Suc-Ala-Ala-Pro-Phe-pNA, to the human cyclophilin hCyp-18 was investigated. HCyp-18 is able to bind 2^4-mer peptides as well as shorter para-nitroaniline (pNA) derivatives and pNA surrogates. Although Suc-Ala-Phe-pNA binds hCyp-18, only proline-containing peptides are able to block efficiently the peptidyl-prolyl cis/trans isomerase activity. Competition experiments strongly suggest the existence of two independent subsites: a S1P P`proline' subsite and a S2P P^S3P P`pNA' subsite. The interaction at S2'^S3' requires either a Phe-pNA C-terminus or a Phe-pNA surrogate bearing an H-bond acceptor able to bind Trp121 and Arg148 simultaneously. ß 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

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Mapping the stereospecificity of peptidyl prolyl cis/trans isomerases

Cited by 31 publications

References 46 publications

Substrate-Based Design of Reversible Pin1 Inhibitors

Substrate-Based Design of Reversible Pin1 Inhibitors

Peptidylprolyl Cis / Trans Isomerases (Immunophilins): Biological Diversity - Targets - Functions

Interaction of human cyclophilin hCyp‐18 with short peptides suggests the existence of two functionally independent subsites

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