LIM domains of cysteine-rich protein 1 (CRP1) are essential for its zyxin-binding function.
Previous studies have demonstrated that the adhesion-plaque protein, zyxin, interacts specifically with a 23 kDa protein, called the cysteine-rich protein 1 (CRP1), which has been implicated in myogenesis. Primary sequence analyses have revealed that both zyxin and CRP1 exhibit multiple copies of a structural motif called the LIM domain. LIM domains, which are defined by the consensus CX2CX16-23HX2CX2CX2CX16-23CX2-3(C,H,D), are found in a variety of proteins that are involved in cell growth and differentiation. Recent studies have established that LIM domains are zinc-binding structures that mediate specific protein-protein interactions. For example, in the case of the zyxin-CRP1 interaction, one of zyxin's three LIM domains is necessary and sufficient for binding to CRP1. Because the CRP1 molecule is comprised primarily of two LIM domains, we were interested in the possibility that the binding site for zyxin on CRP1 might also be contained within a single LIM domain. Consistent with the hypothesis that the LIM domains of CRP1 are critical for the protein's zyxin-binding function, zinc-depleted CRP1 displays a reduced zyxin-binding activity. However, domain mapping analyses have revealed that neither of the two individual LIM domains of CRP1 can support a wild-type interaction with zyxin. Collectively, our results suggest that the binding site for zyxin on CRP1 is not contained within a single contiguous sequence of amino acids. Instead, the interaction appears to rely on the co-ordinate action of a number of residues that are displayed in both of CRP1's LIM domains.
Schmeichel
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Ernest Orlando Lawrence Berkeley National Laboratory, One Cyclotron Road, MS 83-101, Berkeley, CA 94720, USA.
Beckerle
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D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D030161:Avian Proteins; D001665:Binding Sites / Q000502:physiology; D002352:Carrier Proteins / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology; D002645:Chickens; D060588:LIM Domain Proteins; D008667:Metalloproteins / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D017384:Sequence Deletion / Q000235:genetics; D015032:Zinc / Q000172:deficiency / Q000378:metabolism; D016335:Zinc Fingers / Q000502:physiology; D060589:Zyxin
D048868:Adaptor Proteins, Signal Transducing; D030161:Avian Proteins; D002352:Carrier Proteins; D060588:LIM Domain Proteins; D008667:Metalloproteins; D011994:Recombinant Proteins; D060589:Zyxin; D015032:Zinc