TY - JOUR
T1 - Evidence that HAX-1 is an interleukin-1α N-terminal binding protein
AU - Yin, Huali
AU - Morioka, Hideo
AU - Towle, Christine A.
AU - Vidal, Marc
AU - Watanabe, Takeshi
AU - Weissbach, Lawrence
N1 - Funding Information:
We are indebted to Drs Dennis Burke and Henry Mankin for providing osteoarthritic cartilage tissue, and to Marianne Wright and Carol Trahan for technical assistance. We are grateful to Khoa Tran and David Herion for assistance during the early phase of this project, and wish to thank Andre Bernards for supplying the fetal liver cDNA library. We are indebted to Dr Peter Thumfort for suggesting the use of peptide array blots for analysing protein–protein interactions, and for helping design the peptide array. We wish to thank the MIT CCR HHMI Biopolymers Laboratory for synthesis of the oligopeptide array blot. We are grateful to Sam Riley of the MGH Photography Department for guidance with computer graphics. This work was funded in part by grant No. 99-020 from the Orthopaedic Research and Education Foundation, and by an endowment from Dr Harry Wechsler.
PY - 2001/8/7
Y1 - 2001/8/7
N2 - During studies aimed at understanding the function of the N-terminal peptide of interleukin-1α (IL-1 NTP, amino acids 1-112), which is liberated from the remainder of IL-1α during intracellular processing, we identified by yeast two-hybrid analysis a putative interacting protein previously designated as HAX-1. In vitro binding studies and transient transfection experiments confirmed that HAX-1 can associate with the IL-1 NTP. HAX-1 was first identified as a protein that associates with HS1, a target of non-receptor protein tyrosine kinases within haematopoietic cells. Recent data have also revealed interactions between HAX-1 and three disparate proteins, polycystin-2 (derived from the PKD2 gene), a protein linked to polycystic kidney disease, cortactin, and Epstein-Barr virus nuclear antigen leader protein (EBNA-LP). Sequence analysis of different HAX-1 binding domains revealed a putative consensus binding motif that is present in various intracellular proteins. Overlapping peptides comprising the IL-1 NTP were synthesized, and binding experiments revealed that discrete peptides were capable of interacting with HAX-1. HAX-1 may serve to retain the IL-1 NTP in the cytoplasm, and complex formation between the IL-1 NTP and HAX-1 may play a role in motility and/or adhesion of cells.
AB - During studies aimed at understanding the function of the N-terminal peptide of interleukin-1α (IL-1 NTP, amino acids 1-112), which is liberated from the remainder of IL-1α during intracellular processing, we identified by yeast two-hybrid analysis a putative interacting protein previously designated as HAX-1. In vitro binding studies and transient transfection experiments confirmed that HAX-1 can associate with the IL-1 NTP. HAX-1 was first identified as a protein that associates with HS1, a target of non-receptor protein tyrosine kinases within haematopoietic cells. Recent data have also revealed interactions between HAX-1 and three disparate proteins, polycystin-2 (derived from the PKD2 gene), a protein linked to polycystic kidney disease, cortactin, and Epstein-Barr virus nuclear antigen leader protein (EBNA-LP). Sequence analysis of different HAX-1 binding domains revealed a putative consensus binding motif that is present in various intracellular proteins. Overlapping peptides comprising the IL-1 NTP were synthesized, and binding experiments revealed that discrete peptides were capable of interacting with HAX-1. HAX-1 may serve to retain the IL-1 NTP in the cytoplasm, and complex formation between the IL-1 NTP and HAX-1 may play a role in motility and/or adhesion of cells.
KW - HAX-1
KW - In vitro binding
KW - Interleukin-1α
KW - Intracrine signalling
KW - Yeast two-hybrid
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U2 - 10.1006/cyto.2001.0891
DO - 10.1006/cyto.2001.0891
M3 - Article
C2 - 11554782
AN - SCOPUS:0035822817
SN - 1043-4666
VL - 15
SP - 122
EP - 137
JO - Cytokine
JF - Cytokine
IS - 3
ER -