ABCMdb

PMID: 9867828

Propper C, Huang X, Roth J, Sorg C, Nacken W
Analysis of the MRP8-MRP14 protein-protein interaction by the two-hybrid system suggests a prominent role of the C-terminal domain of S100 proteins in dimer formation.
J Biol Chem. 1999 Jan 1;274(1):183-8., [PubMed]

Sentences

No. Mutations Sentence Comment

137 ABCC11 p.Leu95Ala Amino Acid Exchanges of F89A, L95A, and F20A, but Not I17A Abolish Dimerization-Recent publications investigated the dimeric structure of S100 protein calcyclin via NMR spectroscopy (6). Login to comment 138 ABCC11 p.Leu95Ala Amino Acid Exchanges of F89A, L95A, and F20A, but Not I17A Abolish Dimerization-Recent publications investigated the dimeric structure of S100 protein calcyclin via NMR spectroscopy (6). Login to comment 139 ABCC11 p.Leu95Ala To investigate the functional relevance of these residues several murine MRP14 mutants were constructed by exchanging single amino acids (Fig. 2): phenylalanine at position 89 to alanine (F89A), leucine at position 95 to alanine (I95A), isoleucine at position 17 to alanine (I17A), and phenylalanine at position 20 to alanine (F20A). Login to comment 140 ABCC11 p.Leu95Ala To investigate the functional relevance of these residues several murine MRP14 mutants were constructed by exchanging single amino acids (Fig. 2): phenylalanine at position 89 to alanine (F89A), leucine at position 95 to alanine (I95A), isoleucine at position 17 to alanine (I17A), and phenylalanine at position 20 to alanine (F20A). Login to comment 194 ABCC11 p.Leu95Ala Vector encoding GAL4-DNA-BD Vector encoding GAL4-AD mMRP8 mMRP14 beta-GAL units % beta-GAL units % mMRP14 92 Ϯ 20 100 Ϯ 22 28 Ϯ 10 30 Ϯ 11 mMRP8 32 Ϯ 9 35 Ϯ 10 109 Ϯ 17 118 Ϯ 18 mMRP14 (1-101) 37 Ϯ 12 40 Ϯ 13 14 Ϯ 4 15 Ϯ 4 mMRP14 (1-85) Ͻ1 Ͻ1 Ͻ1 Ͻ1 mMRP14 (1-69) Ͻ1 Ͻ1 Ͻ1 Ͻ1 mMRP14 (9-113) 20 Ϯ 9 22 Ϯ 10 12 Ϯ 3 13 Ϯ 3 mMRP14-I17A 23 Ϯ 8 25 Ϯ 9 6 Ϯ 2 7 Ϯ 2 mMRP14-F20A 4 Ϯ 1 4 Ϯ 1 Ͻ1 Ͻ1 mMRP14-F89A 2 Ϯ 1 2 Ϯ 1 Ͻ1 Ͻ1 mMRP14-L95A Ͻ1 Ͻ1 Ͻ1 Ͻ1 mhMRP14 2 Ϯ 1 2 Ϯ 1 Ͻ1 Ͻ1 hmMRP14 64 Ϯ 10 70 Ϯ 11 16 Ϯ 4 17 Ϯ 4 S100A12mMRP14 2 Ϯ 1 2 Ϯ 1 Ͻ1 Ͻ1 S100A12 Ͻ1 Ͻ1 Ͻ1 Ͻ1 Vector encoding GAL4-AD hMRP8 hMRP14 beta-GAL units % beta-GAL units % hMRP14 84 Ϯ 13 100 Ϯ 15 4 Ϯ 1 5 Ϯ 1 hMRP8 7 Ϯ 4 8 Ϯ 5 89 Ϯ 8 106 Ϯ 10 hMRP14-C3S 18 Ϯ 6 21 Ϯ 7 2 Ϯ 1 2 Ϯ 1 hMRP14 (5-114) 16 Ϯ 5 19 Ϯ 6 2 Ϯ 1 2 Ϯ 1 mhMRP14 52 Ϯ 8 58 Ϯ 9 2 Ϯ 1 2 Ϯ 1 hmMRP14 2 Ϯ 1 2 Ϯ 1 3 Ϯ 1 4 Ϯ 1 S100A12mMRP14 Ͻ1 Ͻ1 Ͻ1 Ͻ1 S100A12 Ͻ1 Ͻ1 Ͻ1 Ͻ1 The chimeric human S100A12/murine MRP14 molecule did not dimerize with MRPs in yeast, suggesting that not any N-terminal S100 domain can substitute the homologous one. Login to comment 195 ABCC11 p.Leu95Ala Vector encoding GAL4-DNA-BD Vector encoding GAL4-AD mMRP8 mMRP14 b-GAL units % b-GAL units % mMRP14 92 6 20 100 6 22 28 6 10 30 6 11 mMRP8 32 6 9 35 6 10 109 6 17 118 6 18 mMRP14 (1-101) 37 6 12 40 6 13 14 6 4 15 6 4 mMRP14 (1-85) ,1 ,1 ,1 ,1 mMRP14 (1-69) ,1 ,1 ,1 ,1 mMRP14 (9-113) 20 6 9 22 6 10 12 6 3 13 6 3 mMRP14-I17A 23 6 8 25 6 9 6 6 2 7 6 2 mMRP14-F20A 4 6 1 4 6 1 ,1 ,1 mMRP14-F89A 2 6 1 2 6 1 ,1 ,1 mMRP14-L95A ,1 ,1 ,1 ,1 mhMRP14 2 6 1 2 6 1 ,1 ,1 hmMRP14 64 6 10 70 6 11 16 6 4 17 6 4 S100A12mMRP14 2 6 1 2 6 1 ,1 ,1 S100A12 ,1 ,1 ,1 ,1 Vector encoding GAL4-AD hMRP8 hMRP14 b-GAL units % b-GAL units % hMRP14 84 6 13 100 6 15 4 6 1 5 6 1 hMRP8 7 6 4 8 6 5 89 6 8 106 6 10 hMRP14-C3S 18 6 6 21 6 7 2 6 1 2 6 1 hMRP14 (5-114) 16 6 5 19 6 6 2 6 1 2 6 1 mhMRP14 52 6 8 58 6 9 2 6 1 2 6 1 hmMRP14 2 6 1 2 6 1 3 6 1 4 6 1 S100A12mMRP14 ,1 ,1 ,1 ,1 S100A12 ,1 ,1 ,1 ,1 MRP8-MRP14 Interaction Analyzed by the Two-hybrid System The chimeric human S100A12/murine MRP14 molecule did not dimerize with MRPs in yeast, suggesting that not any N-terminal S100 domain can substitute the homologous one. Login to comment 197 ABCC11 p.Leu95Ala The analysis of three of these mutants, F20A, F89A, and L95A, seems to confirm the importance of these residues. Login to comment 198 ABCC11 p.Leu95Ala The analysis of three of these mutants, F20A, F89A, and L95A, seems to confirm the importance of these residues. Login to comment