Functional and Complementary Phosphorylation State Attributes of Human Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) Isoforms Resolved by Free Flow Electrophoresis

Fetal growth restriction (FGR) is a common disorder in which a fetus is unable to achieve its genetically determined potential size. High concentrations of insulin-like growth factor-binding protein-1 (IGFBP-1) have been associated with FGR. Phosphorylation of IGFBP-1 is a mechanism by which insulin-like growth factor-I (IGF-I) bioavailability can be modulated in FGR. In this study a novel strategy was designed to determine a link between IGF-I affinity and the concomitant phosphorylation state characteristics of IGFBP-1 phosphoisoforms. Using free flow electrophoresis (FFE), multiple IGFBP-1 phosphoisoforms in amniotic fluid were resolved within pH 4.43–5.09. The binding of IGFBP-1 for IGF-I in was determined with The IGF-I affinity for IGFBP-1 between and phosphorylation and of which was the IGF-I binding affinity for IGFBP-1 phosphoisoforms the with for and a was with phosphorylation a in affinity of IGFBP-1 for In of IGFBP-1 phosphoisoforms was between and potential study a for of this novel in the between phosphorylation of IGFBP-1 and FGR. strategy be to with and Fetal growth restriction (FGR) is a common disorder in which a fetus is unable to achieve its genetically determined potential size. High concentrations of insulin-like growth factor-binding protein-1 (IGFBP-1) have been associated with FGR. Phosphorylation of IGFBP-1 is a mechanism by which insulin-like growth factor-I (IGF-I) bioavailability can be modulated in FGR. In this study a novel strategy was designed to determine a link between IGF-I affinity and the concomitant phosphorylation state characteristics of IGFBP-1 phosphoisoforms. Using free flow electrophoresis (FFE), multiple IGFBP-1 phosphoisoforms in amniotic fluid were resolved within pH 4.43–5.09. The binding of IGFBP-1 for IGF-I in was determined with The IGF-I affinity for IGFBP-1 between and phosphorylation and of which was the IGF-I binding affinity for IGFBP-1 phosphoisoforms the with for and a was with phosphorylation a in affinity of IGFBP-1 for In of IGFBP-1 phosphoisoforms was between and potential study a for of this novel in the between phosphorylation of IGFBP-1 and FGR. strategy be to with and The insulin-like growth growth growth flow growth factor-binding growth growth flow growth factor-binding in growth and growth factor-binding protein-1 (IGFBP-1) is a in amniotic fluid of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in growth factor-binding in and and The of IGFBP-1 is to be its phosphorylation Phosphorylation of insulin-like growth binding of bioavailability in insulin-like growth protein-1 (IGFBP-1) IGFBP-1 by Phosphorylation of IGFBP-1 its affinity for IGF-I Phosphorylation of insulin-like growth in and in affinity for IGFBP-1 the of IGF-I with to and growth of bioavailability in insulin-like growth binding is in is a and and the of the fetus growth and binding and amniotic fluid associated with Fetal growth restriction (FGR) is a in which a fetus is unable to achieve its genetically determined potential size. The of IGFBP-1 is in growth and binding in the and fetus and with and of and of binding to growth factor-binding protein-1 (IGFBP-1) growth and of IGFBP-1 have been in and in of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and have the of IGFBP-1 of to concentrations of IGFBP-1 phosphoisoforms with in of the insulin-like growth protein-1 (IGFBP-1) in and its in the of of phosphorylation of insulin-like growth binding protein-1 in of insulin-like growth (IGF-I) and and protein-1 in by growth binding phosphoisoforms of and growth in The of insulin-like growth binding protein-1 phosphoisoforms in with by phosphorylation of IGFBP-1 been to be the of IGFBP-1 phosphorylation in is The in of of IGFBP-1 phosphorylation by in of of insulin-like growth binding phosphoisoforms have been of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and in of the for binding of IGFBP-1 phosphoisoforms Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to of the amniotic insulin-like growth factor-binding and of the insulin-like growth factor-binding protein-1 in is is to a of the and of IGFBP-1 phosphorylation in and free flow electrophoresis of a and to of the state of phosphorylation and IGF-I binding of IGFBP-1 in a a to of this to determine of IGFBP-1 phosphorylation in and be to of IGF-I affinity in the were pH and pH and were for electrophoresis and were of electrophoresis was a the of and IGF-I was was was was The for IGFBP-1 was and was for was was the in was a for which was in this study and for which was the the of in in with growth were a of in was to the of were and for was and to to IGFBP-1 in was a for by to the The was for The was and in pH of the was by the pH by for The was and the of IGFBP-1 to the was was determined the were in the a a of in the was the for the pH of the pH for a pH a and In the of the the in in to the to the pH The was to the In the was and the was The flow for the were and for the the was with a pH was to the pH of and the of was in the and the was the for was with a The was to a with a of was for a of pH to The in to for the were and for and to a of of In the of was to to in the was to of the the were with of pH of the between and the were for by The IGFBP-1 concentrations in to were by of insulin-like growth binding to pH were for of IGFBP-1 by and in was by and by of IGFBP-1 by were In for was a and was the were in in with growth and IGFBP-1 were IGFBP-1 were were for The and were for of and was for The phosphorylation state of IGFBP-1 phosphoisoforms was by to The were by of IGFBP-1 for IGF-I binding was the were The were and and and of IGFBP-1 in the were for the pH were for IGFBP-1 was were by and the Using of IGFBP-1 with IGF-I was determined the with flow was to the to the the flow was of the was of pH were of The was for The were in a of were and concentrations of (IGFBP-1) were for a in and binding were for by the of the the were for the and the the to the binding to The of with of IGFBP-1 Using were and with of IGFBP-1 were and with pH The and were by of with by were with and with were and of IGFBP-1 Using in the were In the with was in of and with of for a The and were to a to which a of was its to a The was in a and for The were of and in The were of pH the of was to the The were and in in in in to by IGFBP-1 to were a with a a and with a flow of was a and the of in and the IGFBP-1 were to The for were and the were The of the in the were for the of the of the in the were to the in with The were for for were and to was was to was to and the was to was a and of and phosphorylation of were a In of were in the of was for a for were to the of were phosphorylation were of IGFBP-1 of IGF-I is in FGR. The for of IGFBP-1 phosphorylation in its with FGR. a novel of IGF-I binding and phosphorylation state characteristics of IGFBP-1 phosphoisoforms. to the strategy was the of IGFBP-1 phosphoisoforms IGF-I affinity of IGFBP-1 in with for for in the affinity of IGFBP-1 for for the in of IGFBP-1 phosphoisoforms between and novel strategy IGFBP-1 phosphoisoforms of the and of IGFBP-1 have been in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and In this for a novel for and IGFBP-1 in a with of and and and in of in Using IGFBP-1 phosphoisoforms with in a for in IGFBP-1 phosphoisoforms were IGFBP-1 of were of IGF-I binding for IGFBP-1 phosphoisoforms was a a mechanism in in phosphorylation of the by a of In phosphorylation of IGFBP-1 and IGFBP-1 is of of the of phosphorylation in the of In the of a of in the of and of in is is a IGFBP-1 is in IGF-I binding IGFBP-1 is of of the Using IGFBP-1 by phosphorylation multiple to be with IGF-I a was in and insulin-like growth binding protein-1 and its phosphorylation of be the phosphorylation associated with affinity for IGF-I in amniotic fluid is and phosphorylation of IGFBP-1 IGF-I binding affinity Phosphorylation of insulin-like growth binding The of the of IGFBP-1 been for the of insulin-like growth by insulin-like growth factor-binding and of the of insulin-like growth factor-binding protein-1 amniotic study a in the of the be by of IGFBP-1 IGF-I for of the multiple phosphorylation and of a have the of IGFBP-1 to in growth and binding in the and fetus and with and of and of binding to growth factor-binding protein-1 (IGFBP-1) growth and The of in the phosphorylation state of IGFBP-1 is of bioavailability in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to for the and of IGFBP-1 phosphoisoforms in (FGR) with the The this study a for the phosphorylation and the of phosphorylation to determine the between IGFBP-1 phosphorylation and have IGFBP-1 phosphoisoforms with to IGF-I binding and binding affinity with phosphorylation of IGFBP-1 phosphoisoforms. to study the of IGFBP-1 phosphorylation been study a which be in of IGFBP-1 phosphorylation with in of phosphorylation of insulin-like growth binding protein-1 in of insulin-like growth (IGF-I) and and protein-1 in by growth The of insulin-like growth binding protein-1 phosphoisoforms in with by growth binding phosphoisoforms of IGFBP-1 and growth in The strategy is for and by The insulin-like growth growth growth flow growth factor-binding growth growth flow growth factor-binding in growth and growth factor-binding protein-1 (IGFBP-1) is a in amniotic fluid of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in growth factor-binding in and and The of IGFBP-1 is to be its phosphorylation Phosphorylation of insulin-like growth binding of bioavailability in insulin-like growth protein-1 (IGFBP-1) IGFBP-1 by Phosphorylation of IGFBP-1 its affinity for IGF-I Phosphorylation of insulin-like growth in and in affinity for IGFBP-1 the of IGF-I with to and growth of bioavailability in insulin-like growth binding is in insulin-like growth amniotic fluid growth restriction free flow electrophoresis insulin-like growth factor-binding IGF-I insulin-like growth amniotic fluid growth restriction free flow electrophoresis insulin-like growth factor-binding IGF-I is a and and the of the fetus growth and binding and amniotic fluid associated with Fetal growth restriction (FGR) is a in which a fetus is unable to achieve its genetically determined potential size. The of IGFBP-1 is in growth and binding in the and fetus and with and of and of binding to growth factor-binding protein-1 (IGFBP-1) growth and of IGFBP-1 have been in and in of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and have the of IGFBP-1 of to concentrations of IGFBP-1 phosphoisoforms with in of the insulin-like growth protein-1 (IGFBP-1) in and its in the of of phosphorylation of insulin-like growth binding protein-1 in of insulin-like growth (IGF-I) and and protein-1 in by growth binding phosphoisoforms of and growth in The of insulin-like growth binding protein-1 phosphoisoforms in with by phosphorylation of IGFBP-1 been to be the of IGFBP-1 phosphorylation in is The in of of IGFBP-1 phosphorylation by in of of insulin-like growth binding IGFBP-1 phosphoisoforms have been of insulin-like growth factor-binding protein-1 (IGFBP-1) phosphoisoforms in and the of IGFBP-1 in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and in of the for binding of IGFBP-1 phosphoisoforms Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to of the amniotic insulin-like growth factor-binding and of the insulin-like growth factor-binding protein-1 in is is to a of the and of IGFBP-1 phosphorylation in and free flow electrophoresis of a and to of the state of phosphorylation and IGF-I binding of IGFBP-1 in a a to of this to determine of IGFBP-1 phosphorylation in and be to of IGF-I affinity in the were pH and pH and were for electrophoresis and were of electrophoresis was a the of and IGF-I was was was was The for IGFBP-1 was and was for was was the in was a for which was in this study and for which was the the of in in with growth were a of in was to the of were and for was and to to IGFBP-1 in was a for by to the The was for The was and in pH of the was by the pH by for The was and the of IGFBP-1 to the was was determined the were in the a a of in the was the for the pH of the pH for a pH a and In the of the the in in to the to the pH The was to the In the was and the was The flow for the were and for the the was with a pH was to the pH of and the of was in the and the was the for was with a The was to a with a of was for a of pH to The in to for the were and for and to a of of In the of was to to in the was to of the the were with of pH of the between and the were for by The IGFBP-1 concentrations in to were by of insulin-like growth binding to pH were for of IGFBP-1 by and in was by and by of IGFBP-1 by were In for was a and was the were in in with growth and IGFBP-1 were IGFBP-1 were were for The and were for of and was for The phosphorylation state of IGFBP-1 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phosphorylation were were and with of IGFBP-1 were and with pH The and were by of with by were with and with were and of IGFBP-1 Using in the were In the with was in of and with of for a The and were to a to which a of was its to a The was in a and for The were of and in The were of pH the of was to the The were and in in in in to by IGFBP-1 to were a with a a and with a flow of was a and the of in and the IGFBP-1 were to The for were and the were The of the in the were for the of the of the in the were to the in with The were for for were and to was was to was to and the was to was a and of and phosphorylation of were a In of were in the of was for a for were to the of were phosphorylation were were and with of IGFBP-1 were and with pH The and were by of with by were with and with were and The were and with of IGFBP-1 were and with pH The and were by of with by were with and with were and of IGFBP-1 Using in the were In the with was in of and with of for a The and were to a to which a 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The for of IGFBP-1 phosphorylation in its with FGR. a novel of IGF-I binding and phosphorylation state characteristics of IGFBP-1 phosphoisoforms. to the strategy was the of IGFBP-1 phosphoisoforms IGF-I affinity of IGFBP-1 in with for for in the affinity of IGFBP-1 for for the in of IGFBP-1 phosphoisoforms between and novel strategy IGFBP-1 phosphoisoforms of the and of IGFBP-1 have been in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and In this for a novel for and IGFBP-1 in a with of and and and in of in Using IGFBP-1 phosphoisoforms with in a for in IGFBP-1 phosphoisoforms were IGFBP-1 of were of IGF-I binding for IGFBP-1 phosphoisoforms was a a mechanism in in phosphorylation of the by a of In phosphorylation of IGFBP-1 and IGFBP-1 is of of the of phosphorylation in the of In the of a of in the of and of in is is a IGFBP-1 is in IGF-I binding IGFBP-1 is of of the Using IGFBP-1 by phosphorylation multiple to be with IGF-I a was in and insulin-like growth binding protein-1 and its phosphorylation of be the phosphorylation associated with affinity for IGF-I in amniotic fluid is and phosphorylation of IGFBP-1 IGF-I binding affinity Phosphorylation of insulin-like growth binding The of the of IGFBP-1 been for the of insulin-like growth by insulin-like growth factor-binding and of the of insulin-like growth factor-binding protein-1 amniotic study a in the of the be by of IGFBP-1 IGF-I for of the multiple phosphorylation and of a have the of IGFBP-1 to in growth and binding in the and fetus and with and of and of binding to growth factor-binding protein-1 (IGFBP-1) growth and The of in the phosphorylation state of IGFBP-1 is of bioavailability in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to for the and of IGFBP-1 phosphoisoforms in (FGR) with the The this study a for the phosphorylation and the of phosphorylation to determine the between IGFBP-1 phosphorylation and have IGFBP-1 phosphoisoforms with to IGF-I binding and binding affinity with phosphorylation of IGFBP-1 phosphoisoforms. to study the of IGFBP-1 phosphorylation been study a which be in of IGFBP-1 phosphorylation with in of phosphorylation of insulin-like growth binding protein-1 in of insulin-like growth (IGF-I) and and protein-1 in by growth The of insulin-like growth binding protein-1 phosphoisoforms in with by growth binding phosphoisoforms of IGFBP-1 and growth in The strategy is for and by Phosphorylation of IGFBP-1 of IGF-I is in FGR. The for of IGFBP-1 phosphorylation in its with FGR. a novel of IGF-I binding and phosphorylation state characteristics of IGFBP-1 phosphoisoforms. to the strategy was the of IGFBP-1 phosphoisoforms IGF-I affinity of IGFBP-1 in with for for in the affinity of IGFBP-1 for for the in of IGFBP-1 phosphoisoforms between and novel strategy IGFBP-1 phosphoisoforms of the and of IGFBP-1 have been in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to The phosphorylation of insulin-like growth factor-binding protein-1 in is in amniotic fluid and In this for a novel for and IGFBP-1 in a with of and and and in of in Using IGFBP-1 phosphoisoforms with in a for in IGFBP-1 phosphoisoforms were IGFBP-1 of were of IGF-I binding for IGFBP-1 phosphoisoforms was a a mechanism in in phosphorylation of the by a of In phosphorylation of IGFBP-1 and IGFBP-1 is of of the of phosphorylation in the of In the of a of in the of and of in is is a IGFBP-1 is in IGF-I binding IGFBP-1 is of of the Using IGFBP-1 by phosphorylation multiple to be with IGF-I a was in and insulin-like growth binding protein-1 and its phosphorylation of be the phosphorylation associated with affinity for IGF-I in amniotic fluid is and phosphorylation of IGFBP-1 IGF-I binding affinity Phosphorylation of insulin-like growth binding The of the of IGFBP-1 been for the of insulin-like growth by insulin-like growth factor-binding and of the of insulin-like growth factor-binding protein-1 amniotic study a in the of the be by of IGFBP-1 IGF-I for of the multiple phosphorylation and of a have the of IGFBP-1 to in growth and binding in the and fetus and with and of and of binding to growth factor-binding protein-1 (IGFBP-1) growth and The of in the phosphorylation state of IGFBP-1 is of bioavailability in Phosphorylation of insulin-like growth factor-binding protein-1 in amniotic fluid and to for the and of IGFBP-1 phosphoisoforms in (FGR) with the The this study a for the phosphorylation and the of phosphorylation to determine the between IGFBP-1 phosphorylation and FGR. In have IGFBP-1 phosphoisoforms with to IGF-I binding and binding affinity with phosphorylation of IGFBP-1 phosphoisoforms. to study the of IGFBP-1 phosphorylation been study a which be in of IGFBP-1 phosphorylation with in of phosphorylation of insulin-like growth binding protein-1 in of insulin-like growth (IGF-I) and and protein-1 in by growth The of insulin-like growth binding protein-1 phosphoisoforms in with by growth binding phosphoisoforms of IGFBP-1 and growth in The strategy is for and by of for and the for and in to for and in the of a and of a in

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