Chaperone‐like activity of peptidyl‐prolyl cis‐trans isomerase during creatine kinase refolding

Abstract

Porcine kidney 18 kD peptidyl-prolyl cis-trans isomerase (PPIase) belongs to the cyclophilin family that is inhibited by the immunosuppressive drug cyclosporin A. The chaperone activity of PPIase was studied using inactive, active, and alkylated PPIase during rabbit muscle creatine kinase (CK) refolding. The results showed that low concentration inactive or active PPIase was able to improve the refolding yields, while high concentration PPIase decreased the CK reactivation yields. Aggregation was inhibited by inactive or active PPIase, and completely suppressed at 32 or 80 times the CK concentration (2.7 M). However, alkylated PPIase was not able to prevent CK aggregation. In addition, the ability of inactive PPIase to affect CK reactivation and prevent CK aggregation was weaker than that of active PPIase. These results indicate that PPIase interacted with the early folding intermediates of CK, thus preventing their aggregation in a concentration-dependent manner. PPIase exhibited chaperone-like activity during CK refolding. The results also suggest that the isomerase activity of PPIase was independent of the chaperone activity, and that the proper molar ratio was important for the chaperone activity of PPIase. The cysteine residues of PPIase may be a peptide binding site, and may be an essential group for the chaperone function.

Catalysis of proline isomerization and molecular chaperone activity in a tug-of-war

“…The opposing effects are in agreement with findings that CypA has a PPIase-independent, general chaperone activity that is important for leukocyte chemotaxis 44 and in vitro refolding of arginine kinase 45 . In addition, previous studies have shown opposing effects of CypA on protein folding at low and high CypA concentrations 46 .…”

Catalysis of proline isomerization and molecular chaperone activity in a tug-of-war

“…The opposing effects are in agreement with findings that CypA has a PPIase-independent, general chaperone activity that is important for leukocyte chemotaxis 44 and in vitro refolding of arginine kinase 45 . In addition, previous studies have shown opposing effects of CypA on protein folding at low and high CypA concentrations 46 .…”

Comparative Physiological and Proteomic Analysis Reveal Distinct Regulation of Peach Skin Quality Traits by Altitude

“…The induction of these HSPs (also called “molecular chaperones”) along with the up-regulation of peptidyl-prolyl cis-trans isomerase (Table 1), that also act as chaperone (Ou et al, 2001), is interesting because it was suggested that chaperones act as shields protecting proteins against oxidative damage, notably during peach fruit ripening (Nilo et al, 2010). In addition, it is thought that chaperones closely interact with misfolded or damaged proteins to assist their refolding upon environmental stress (Magi et al, 2004).…”

Designing an Antibody-Based Chaperoning System through Programming the Binding and Release of the Folding Intermediate

“…Ou et al studied the chaperone-like effect of porcine kidney 18 kDa peptidyl-prolyl cis–trans isomerase (Cyp18) on folding of rabbit muscle CK and found Cyp18 could inhibit protein aggregation. They also found Cyp18 at low concentrations was able to improve the refolding yields of CK (from 70 to 88%), while a high concentration of Cyp18 caused a decrease in refolding yields . Zhao et al studied the effect of protein disulfide isomerase (PDI) on folding of rabbit muscle CK.…”

Catalysis of proline isomerization and molecular chaperone activity in a tug-of-war

“…The opposing effects are in agreement with findings that CypA has a PPIase-independent, general chaperone activity that is important for leukocyte chemotaxis 44 and in vitro refolding of arginine kinase 45 . In addition, previous studies have shown opposing effects of CypA on protein folding at low and high CypA concentrations 46 .…”

Comparative Physiological and Proteomic Analysis Reveal Distinct Regulation of Peach Skin Quality Traits by Altitude

“…The induction of these HSPs (also called “molecular chaperones”) along with the up-regulation of peptidyl-prolyl cis-trans isomerase (Table 1), that also act as chaperone (Ou et al, 2001), is interesting because it was suggested that chaperones act as shields protecting proteins against oxidative damage, notably during peach fruit ripening (Nilo et al, 2010). In addition, it is thought that chaperones closely interact with misfolded or damaged proteins to assist their refolding upon environmental stress (Magi et al, 2004).…”

Designing an Antibody-Based Chaperoning System through Programming the Binding and Release of the Folding Intermediate

“…Ou et al studied the chaperone-like effect of porcine kidney 18 kDa peptidyl-prolyl cis–trans isomerase (Cyp18) on folding of rabbit muscle CK and found Cyp18 could inhibit protein aggregation. They also found Cyp18 at low concentrations was able to improve the refolding yields of CK (from 70 to 88%), while a high concentration of Cyp18 caused a decrease in refolding yields . Zhao et al studied the effect of protein disulfide isomerase (PDI) on folding of rabbit muscle CK.…”

Catalysis of proline isomerization and molecular chaperone activity in a tug-of-war

“…The opposing effects are in agreement with findings that CypA has a PPIase-independent, general chaperone activity that is important for leukocyte chemotaxis 44 and in vitro refolding of arginine kinase 45 . In addition, previous studies have shown opposing effects of CypA on protein folding at low and high CypA concentrations 46 .…”

Comparative Physiological and Proteomic Analysis Reveal Distinct Regulation of Peach Skin Quality Traits by Altitude

“…The induction of these HSPs (also called “molecular chaperones”) along with the up-regulation of peptidyl-prolyl cis-trans isomerase (Table 1), that also act as chaperone (Ou et al, 2001), is interesting because it was suggested that chaperones act as shields protecting proteins against oxidative damage, notably during peach fruit ripening (Nilo et al, 2010). In addition, it is thought that chaperones closely interact with misfolded or damaged proteins to assist their refolding upon environmental stress (Magi et al, 2004).…”

Designing an Antibody-Based Chaperoning System through Programming the Binding and Release of the Folding Intermediate

“…Ou et al studied the chaperone-like effect of porcine kidney 18 kDa peptidyl-prolyl cis–trans isomerase (Cyp18) on folding of rabbit muscle CK and found Cyp18 could inhibit protein aggregation. They also found Cyp18 at low concentrations was able to improve the refolding yields of CK (from 70 to 88%), while a high concentration of Cyp18 caused a decrease in refolding yields . Zhao et al studied the effect of protein disulfide isomerase (PDI) on folding of rabbit muscle CK.…”