Expression, purification, crystallization and preliminary X-ray analysis of the olfactomedin domain from the sea urchin cell-adhesion protein amassin.
A family of animal proteins is emerging which contain a conserved protein motif known as an olfactomedin (OLF) domain. Novel extracellular protein-protein interactions occur through this domain. The OLF-family member amassin, from the sea urchin Strongylocentrotus purpuratus, has previously been identified to mediate a rapid cell-adhesion event resulting in a large aggregation of coelomocytes, the circulating immune cells. In this work, heterologous expression and purification of the OLF domain from amassin was carried out and initial crystallization trials were performed. A native data set has been collected, extending to 2.7 A under preliminary cryoconditions, using an in-house generator. This work leads the way to the determination of the first structure of an OLF domain.
Hillier
Brian J
BJ
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA 92093-0202, USA. bhillier@ucsd.edu
Sundaresan
Vidyasankar
V
Stout
C David
CD
Vacquier
Victor D
VD
D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D047270:Strongylocentrotus purpuratus
D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D011994:Recombinant Proteins; C081451:olfactomedin