Expression Cloning and Biochemical Characterizations
of Recombinant Cyclophilin Proteins from
Schistosoma mansoni

Francesca Bugli; A Khattab; E Vigneti; R Butler; D Cioli; Mq Klinkert

doi:PT970852

Expression Cloning and Biochemical Characterizations of Recombinant Cyclophilin Proteins from Schistosoma mansoni

Francesca Bugli
, A Khattab
, E Vigneti
, R Butler
, D Cioli
, Mq Klinkert

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

Recombinant Schistosoma mansoni cyclophilin proteins of the A and the B subtypes (SmCYP A and B) were expressed in bacterial cells as histidine- and maltose-binding fusion proteins and also as nonfused proteins. In addition, S. mansoni CYPs were produced in Sf9 insect cells in their natural forms. Purified recombinant SmCYP B was found to possess a peptidyl-prolyl cis-trans isomerase (PPIase) activity, with a kcat/Km value of 8.2 x 10(5) M-1 s-1. The SmCYP B isoform is approximately two to three times more active than SmCYP A. SmCYP B-specific RNA appears to be more abundant in adult schistosomes than SmCYP A RNA in Northern blots. These results support the conclusion that SmCYP B represents the major schistosomal CYP. The PPIase-associated activity of both CYPs was inhibitable by the immunosuppressive drug cyclosporin A (CsA). We attempt to explain differences in PPIase activities and in CsA inhibition by examining models of the two CYPs complexed to CsA

Original language	English
Pages (from-to)	240-246
Number of pages	7
Journal	Protein Expression and Purification
Volume	12
DOIs	https://doi.org/PT970852
Publication status	Published - 1998

Keywords

Cyclosporin A
Recombinant Cyclophilin

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PT970852

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@article{9fa393d8e8a74c908b994c48501446c8,

title = "Expression Cloning and Biochemical Characterizations of Recombinant Cyclophilin Proteins from Schistosoma mansoni",

abstract = "Recombinant Schistosoma mansoni cyclophilin proteins of the A and the B subtypes (SmCYP A and B) were expressed in bacterial cells as histidine- and maltose-binding fusion proteins and also as nonfused proteins. In addition, S. mansoni CYPs were produced in Sf9 insect cells in their natural forms. Purified recombinant SmCYP B was found to possess a peptidyl-prolyl cis-trans isomerase (PPIase) activity, with a kcat/Km value of 8.2 x 10(5) M-1 s-1. The SmCYP B isoform is approximately two to three times more active than SmCYP A. SmCYP B-specific RNA appears to be more abundant in adult schistosomes than SmCYP A RNA in Northern blots. These results support the conclusion that SmCYP B represents the major schistosomal CYP. The PPIase-associated activity of both CYPs was inhibitable by the immunosuppressive drug cyclosporin A (CsA). We attempt to explain differences in PPIase activities and in CsA inhibition by examining models of the two CYPs complexed to CsA",

keywords = "Cyclosporin A, Recombinant Cyclophilin, Cyclosporin A, Recombinant Cyclophilin",

author = "Francesca Bugli and A Khattab and E Vigneti and R Butler and D Cioli and Mq Klinkert",

year = "1998",

doi = "PT970852",

language = "English",

volume = "12",

pages = "240--246",

journal = "Protein Expression and Purification",

issn = "1046-5928",

publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Expression Cloning and Biochemical Characterizations of Recombinant Cyclophilin Proteins from Schistosoma mansoni

AU - Bugli, Francesca

AU - Khattab, A

AU - Vigneti, E

AU - Butler, R

AU - Cioli, D

AU - Klinkert, Mq

PY - 1998

Y1 - 1998

N2 - Recombinant Schistosoma mansoni cyclophilin proteins of the A and the B subtypes (SmCYP A and B) were expressed in bacterial cells as histidine- and maltose-binding fusion proteins and also as nonfused proteins. In addition, S. mansoni CYPs were produced in Sf9 insect cells in their natural forms. Purified recombinant SmCYP B was found to possess a peptidyl-prolyl cis-trans isomerase (PPIase) activity, with a kcat/Km value of 8.2 x 10(5) M-1 s-1. The SmCYP B isoform is approximately two to three times more active than SmCYP A. SmCYP B-specific RNA appears to be more abundant in adult schistosomes than SmCYP A RNA in Northern blots. These results support the conclusion that SmCYP B represents the major schistosomal CYP. The PPIase-associated activity of both CYPs was inhibitable by the immunosuppressive drug cyclosporin A (CsA). We attempt to explain differences in PPIase activities and in CsA inhibition by examining models of the two CYPs complexed to CsA

AB - Recombinant Schistosoma mansoni cyclophilin proteins of the A and the B subtypes (SmCYP A and B) were expressed in bacterial cells as histidine- and maltose-binding fusion proteins and also as nonfused proteins. In addition, S. mansoni CYPs were produced in Sf9 insect cells in their natural forms. Purified recombinant SmCYP B was found to possess a peptidyl-prolyl cis-trans isomerase (PPIase) activity, with a kcat/Km value of 8.2 x 10(5) M-1 s-1. The SmCYP B isoform is approximately two to three times more active than SmCYP A. SmCYP B-specific RNA appears to be more abundant in adult schistosomes than SmCYP A RNA in Northern blots. These results support the conclusion that SmCYP B represents the major schistosomal CYP. The PPIase-associated activity of both CYPs was inhibitable by the immunosuppressive drug cyclosporin A (CsA). We attempt to explain differences in PPIase activities and in CsA inhibition by examining models of the two CYPs complexed to CsA

KW - Cyclosporin A

KW - Recombinant Cyclophilin

KW - Cyclosporin A

KW - Recombinant Cyclophilin

UR - http://hdl.handle.net/10807/10733

U2 - PT970852

DO - PT970852

M3 - Article

SN - 1046-5928

VL - 12

SP - 240

EP - 246

JO - Protein Expression and Purification

JF - Protein Expression and Purification

ER -

Expression Cloning and Biochemical Characterizations of Recombinant Cyclophilin Proteins from Schistosoma mansoni

Abstract

Keywords

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