A Synergism between Temporins toward Gram-negative Bacteria Overcomes Resistance Imposed by the Lipopolysaccharide Protective Layer

Temporins are short and homologous antimicrobial peptides (AMPs) isolated from the frog skin of Rana genus. To date, very little is known about the biological significance of the presence of closely related AMPs in single living organisms. Here we addressed this question using temporins A, B, and L isolated from Rana temporaria. We found that temporins A and B are only weakly active toward Gram-negative bacteria. However, a marked synergism occurs is To the in we the of the the and of and and the of the of in and using and that L A and B in the the To the of this is the that a of AMPs from the the Temporins are short and homologous antimicrobial peptides (AMPs) isolated from the frog skin of Rana genus. To date, very little is known about the biological significance of the presence of closely related AMPs in single living organisms. Here we addressed this question using temporins A, B, and L isolated from Rana temporaria. We found that temporins A and B are only weakly active toward Gram-negative bacteria. However, a marked synergism occurs is To the in we the of the the and of and and the of the of in and using and that L A and B in the the To the of this is the that a of AMPs from the the of of peptides peptides of the found the living of antimicrobial peptides antimicrobial antimicrobial are of about AMPs in and of and a and the a the of the and However, a AMPs the of and a of AMPs and frog of peptides of closely related and from the and from the frog from the frog and from the Rana and of of AMPs from the and isolated from and and the temporins the of AMPs and in a single are the AMPs found in only a a from and are active toward bacteria. a of this is and Gram-negative biological and of temporins A B and L from Rana temporaria. peptides the of the the of and the in a and significance of the of of AMPs the addressed in only a a synergism bacteria. the and from of and and and isolated from However, the the the is we the synergism of L temporins A B the the of the of Gram-negative a of this is is the of B and is the of AMPs that L the of temporins A and B the the and from and and from in and a using of the peptides from the in To the the from the of the peptides the of the peptides peptides and in the of a the and temporins A B and L from the the peptides a in using a of in a of peptides and and of the the the the using of in a of in of the in in of the a of the the of of of a synergism temporins of in a in a of of L and temporins A and of peptides the A and B are the of A and B in the and are the of A and B and and are the of A and the and isolated from the of the and of the of of the of temporins and temporins A and B, and in and a of in and in and the of of of the from a and in and in a of in the a of the and a of of the of the the the in in and in of Temporins the of and of of a of a of A and and and L and and the peptides the of of the a the of of and in a a in and the peptides a of Temporins the the of a of of temporins A B and L the of and in in the the of the of the in a using and and of the of the peptides A and temporins B and of and in the of the the of of using the and and in and the in that the of the the in in the the peptides we the of temporins A, B, and L using a the in a a of a of peptides peptides in the presence of using of in of the of the of from the of the and the of the of the we a a and peptides in of in a and and the the a of the a of the in the and of the a of the of the the the of the a the and To we the using the of the in the and of the the from the the and the of the and the of the the of a the of the of of the peptides the of the in the of Temporins toward the of of temporins A, B, and L the of Gram-negative and using the are in of and temporins and a of the Gram-negative and the synergism found of and the of temporins A L and B found A B the Gram-negative the peptides from the of of temporins A, B, and of A B A in a of Temporins the the of the of temporins is from of a A marked and of the of temporins A L B L the that L the temporins the of the is are the peptides this in the of the we the of the of and that a only a in of temporins the the of Gram-negative and the that the of the a of of temporins A and L of temporins A and L the of and of in a of of A and and and L and and peptides the of are the of from a single of B, of temporins A and L the of of in in a peptides and in and A L and A L the of from a single of A found B and are of the of in the of the temporins in we the peptides a that is only the of that this the biological of temporins are and the are in the is of the a in the of the a the is that the of A and B, are in and in the of is in of the peptides and the of of the A, B, and a L and the temporins A B the of the in a that of the peptides is the is and of the in A and A and B, a of the the of the of of L in a in that the in the presence of of L the in in of of of the a the and in and is the and the of A, A B, B L the of from a single of of the of the of the of the peptides the of the of temporins A and B We a the and is A the of L and A and B is that temporins A and B are the of in the of the the and the of L the antimicrobial that this the of L Gram-negative in that the the and and the of the of temporins Temporins A B and L in and in and of in and of is the the are the of L of Temporins A and the of the the the of L the of temporins A and B in the presence of We the of and in of the temporins and the of the found A, B, and of A and B the peptides in A B A and that L of temporins A and temporins A and B the of the only of A B and L the of temporins in the presence of A B and L the of the a and in and the of is the the of A and B the the B and A, are the A and B in the the of from a single of Temporins in and in the of synergism in the of we the of in in and peptides the of the peptides the of are the peptides the peptides L a and and temporins A and B in of peptides A L A is in and the in and that the is the of L temporins A and of in and in A, of A B L in B, of A B A L in of the and of the and the of the peptides of a in a from the of the Here we the of the the of the peptides that are are in of the the the a the is a of a of temporins in and A, of the of peptides in the of the in the are the of using a of the are the the the B, the of temporins the of are in the of the of the the are of the of the of of the peptides the of the of the the of this a in the a of the and the peptides a of L A the and of the is in the of L A, the of and antimicrobial toward Gram-negative bacteria. the of L the this from the the and date, very little is known about the biological significance of the presence of closely related AMPs in single living organisms. and the of in frog skin and a of that a of AMPs the a of However, the of this synergism is Temporins of related isolated from skin in Rana of and and are the of AMPs the the of the of the is a this we we the of temporins A and B that are Gram-negative we the a temporins Gram-negative a this of Gram-negative is of the is of the the the peptides and the and the a the found peptides we the of temporins A and B toward Gram-negative bacteria. peptides in the the of of A and B, the of A and is in the active in is in that of AMPs a in antimicrobial Gram-negative the of in A and B the of peptides that this that temporins A and B of the and in the is the that the the is temporins A and B, that L that temporins A and B that of L and that temporins A and B are the the active of the is the of L A B, and the this of is from that the found of the the of the the of the of Gram-negative bacteria. is the in temporins the the of the of a that this synergism is related the of L temporins A and B the a that of and the A and B that a peptides that However, in the presence of the of temporins A and B is only a of in the peptides the of the the peptides are L the of temporins A and B the the To the of this is the peptides the of that the synergism of the of a of is found in of the of in the of B in this is of in in and found and that is peptides of frog skin and in the synergism peptides that the the a and a of the the we that a of and related peptides of antimicrobial in the of the peptides the this of a synergism AMPs from the and a synergism in this the of a of the of the of of peptides peptides of the found the living of antimicrobial peptides antimicrobial antimicrobial are of about AMPs in and of and a and the a the of the and However, a AMPs the of and a of AMPs and frog of peptides of closely related and from the and from the frog from the frog and from the Rana and of of AMPs from the and isolated from and and the temporins the of AMPs and in a single are the AMPs found in only a a from and are active toward bacteria. a of this is and Gram-negative biological and of temporins A B and L from Rana temporaria. peptides the of the the of and the in a and significance of the of of AMPs the addressed in only a a synergism bacteria. the and from of and and and isolated from However, the the the is we the synergism of L temporins A B the the of the of Gram-negative a of this is is the of B and is the of AMPs that L the of temporins A and B the the and from and and from in and a using of the peptides from the in To the the from the of the peptides the of the peptides peptides and in the of a the and temporins A B and L from the the peptides a in using a of in a of peptides and and of the the the the using of in a of in of the in in of the a of the the of of of a synergism temporins of in a in a of of L and temporins A and of peptides the A and B are the of A and B in the and are the of A and B and and are the of A and the and isolated from the of the and of the of of the of temporins and temporins A and B, and in and a of in and in and the of of of the from a and in and in a of in the a of the and a of of the of the the the in in and in of Temporins the of and of of a of a of A and and and L and and the peptides the of of the a the of of and in a a in and the peptides a of Temporins the the of a of of temporins A B and L the of and in in the the of the of the in a using and and of the of the peptides A and temporins B and of and in the of the the of of using the and and in and the in that the of the the in in the the peptides we the of temporins A, B, and L using a the in a a of a of peptides peptides in the presence of using of in of the of the of from the of the and the of the of the we a a and peptides in of in a and and the the a of the a of the in the and of the a of the of the the the of the a the and To we the using the of the in the and of the the from the the and the of the and the of the the of a the of the of of the peptides the of the in the of and from and and from in and a using of the peptides from the in To the the from the of the peptides the of the peptides peptides and in the of a the and temporins A B and L from the the peptides a in using a of in a of peptides and and of the the the the using of in a of in of the in in of the a of the the of of of a synergism temporins of in a in a of of L and temporins A and of peptides the A and B are the of A and B in the and are the of A and B and and are the of A and the and isolated from the of the and of the of of the of temporins and temporins A and B, and in and a of in and in and the of of of the from a and in and in a of in the a of the and a of of the of the the the in in and in of Temporins the of and of of a of a of A and and and L and and the peptides the of of the a the of of and in a a in and the peptides a of Temporins the the of a of of temporins A B and L the of and in in the the of the of the in a using and and of the of the peptides A and temporins B and of and in the of the the of of using the and and in and the in that the of the the in in the the peptides we the of temporins A, B, and L using a the in a a of a of peptides peptides in the presence of using of in of the of the of from the of the and the of the of the we a a and peptides in of in a and and the the a of the a of the in the and of the a of the of the the the of the a the and To we the using the of the in the and of the the from the the and the of the and the of the the of a the of the of of the peptides the of the in the of Temporins toward the of of temporins A, B, and L the of Gram-negative and using the are in of and temporins and a of the Gram-negative and the synergism found of and the of temporins A L and B found A B the Gram-negative the peptides from the of of temporins A, B, and of A B A in a of Temporins the the of the of temporins is from of a A marked and of the of temporins A L B L the that L the temporins the of the is are the peptides this in the of the we the of the of and that a only a in of temporins the the of Gram-negative and the that the of the a of of temporins A and L of the of in the of the temporins in we the peptides a that is only the of that this the biological of temporins are and the are in the is of the a in the of the a the is that the of A and B, are in and in the of is in of the peptides and the of of the A, B, and a L and the temporins A B the of the in a that of the peptides is the is and of the in A and A and B, a of the the of the of of L in a in that the in the presence of of L the in in of of of the a the and in and is the and the of A, A B, B L the of from a single of of the of the of the of the peptides the of the of temporins A and B We a the and is A the of L and A and B is that temporins A and B are the of in the of the the and the of L the antimicrobial that this the of L Gram-negative in that the the and and the of the of temporins Temporins A B and L in and in and of in and of is the the are the of L of Temporins A and the of the the the of L the of temporins A and B in the presence of We the of and in of the temporins and the of the found A, B, and of A and B the peptides in A B A and that L of temporins A and temporins A and B the of the only of A B and L the of temporins in the presence of A B and L the of the a and in and the of is the the of A and B the the B and A, are the A and B in the the of from a single of Temporins in and in the of synergism in the of we the of in in and peptides the of the peptides the of are the peptides the peptides L a and and temporins A and B in of peptides A L A is in and the in and that the is the of L temporins A and of in and in A, of A B L in B, of A B A L in of the and of the and the of the peptides of a in a from the of the Here we the of the the of the peptides that are are in of the the the a the is a of a of temporins in and A, of the of peptides in the of the in the are the of using a of the are the the the B, the of temporins the of are in the of the of the the are of the of the of of the peptides the of the of the the of this a in the a of the and the peptides a of L A the and of the is in the of L A, the of and antimicrobial toward Gram-negative bacteria. the of L the this from the the and Temporins toward the of of temporins A, B, and L the of Gram-negative and using the are in of and temporins and a of the Gram-negative and the synergism found of and the of temporins A L and B found A B the Gram-negative the peptides from the of of Temporins the the of the of temporins is from of a A marked and of the of temporins A L B L the that L the temporins the of the is are the peptides To this in the of the we the of the of and that a only a in of temporins the the of Gram-negative and the that the of the a of of temporins A and L of the of in the of the temporins in we the peptides a that is only the of that this the biological of temporins are and the are in the is of the a in the of the a the is that the of A and B, are in and in the of is in of the peptides and the of of the A, B, and a L and the temporins A B the of the in a that of the peptides is the is and of the in A and A and B, a of the the of the of of L in a in that the in the presence of of L the in in of of the of the of the of the peptides the of the of temporins A and B We a the and is A the of L and A and B is that temporins A and B are the of in the of the the and the of L the antimicrobial that this the of L Gram-negative in that the the and and the L of Temporins A and the of the the the of L the of temporins A and B in the presence of We the of and in of the temporins and the of the found A, B, and of A and B the peptides in A B A and that L of temporins A and temporins A and B the of the only Temporins in and in the of synergism in the of we the of in in and peptides the of the peptides the of are the peptides the peptides L a and and temporins A and B in of peptides A L A is in and the in and that the is the of L temporins A and of the and the of the peptides of a in a from the of the Here we the of the the of the peptides that are are in of the the the a the is a of a of of the peptides the of the of the the of this a in the a of the and the peptides a of L A the and of the is in the of L A, the of and antimicrobial toward Gram-negative bacteria. the of L the this from the the and date, very little is known about the biological significance of the presence of closely related AMPs in single living organisms. and the of in frog skin and a of that a of AMPs the a of However, the of this synergism is Temporins of related isolated from skin in Rana of and and are the of AMPs the the of the of the is a this we we the of temporins A and B that are Gram-negative we the a temporins Gram-negative a this of Gram-negative is of the is of the the the peptides and the and the a the found peptides we the of temporins A and B toward Gram-negative bacteria. peptides in the the of of A and B, the of A and is in the active in is in that of AMPs a in antimicrobial Gram-negative the of in A and B the of peptides that this that temporins A and B of the and in the is the that the the is temporins A and B, that L that temporins A and B that of L and that temporins A and B are the the active of the is the of L A B, and the this of is from that the found of the the of the the of the of Gram-negative bacteria. is the in temporins the the of the of a that this synergism is related the of L temporins A and B the a that of and the A and B that a peptides that However, in the presence of the of temporins A and B is only a of in the peptides the of the the peptides are L the of temporins A and B the the To the of this is the peptides the of that the synergism of the of a of is found in of the of in the of B in this is of in in and found and that is peptides of frog skin and in the synergism peptides that the the a and a of the the we that a of and related peptides of antimicrobial in the of the peptides the this of a synergism AMPs from the and a synergism in this the of a of the of the To date, very little is known about the biological significance of the presence of closely related AMPs in single living organisms. and the of in frog skin and a of that a of AMPs the a of However, the of this synergism is Temporins of related isolated from skin in Rana of and and are the of AMPs the the of the of the is a this we we the of temporins A and B that are Gram-negative we the a temporins Gram-negative a this of Gram-negative is of the is of the the the peptides and the and the a the found peptides we the of temporins A and B toward Gram-negative bacteria. peptides in the the of of A and B, the of A and is in the active in is in that of AMPs a in antimicrobial Gram-negative the of in A and B the of peptides that this that temporins A and B of the and in the is the that the the is temporins A and B, that L that temporins A and B that of L and that temporins A and B are the the active of the is the of L A B, and the this of is from that the found of the the of the the of the of Gram-negative bacteria. is the in temporins the the of the of a that this synergism is related the of L temporins A and B the a that of and the A and B that a peptides that However, in the presence of the of temporins A and B is only a of in the peptides the of the the peptides are L the of temporins A and B the the To the of this is the peptides the of that the synergism of the of a of is found in of the of in the of B in this is of in in and found and that is peptides of frog skin and in the synergism peptides that the the a and a of the the we that a of and related peptides of antimicrobial in the of the peptides the this of a synergism AMPs from the and a synergism in this the of a of the of the We from the of