Linked Life Data

8144540

Source:http://linkedlifedata.com/resource/pubmed/id/8144540

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
13
pubmed:dateCreated
1994-5-5
pubmed:abstractText
The NH2-terminal 29-kDa Fib-1 fragment consisting of the first five finger modules of fibronectin (F1-5) binds reversibly to fibrin and facilitates cross-linking by Factor XIII. To narrow down the fibrin-binding site within this region, we have used recombinant technology to express a number of individual fingers, rF1, rF2, rF3, rF4, and rF5, and their pairs, rF1-2 rF2-3, and rF4-5, as fusion proteins in Escherichia coli. These recombinant fragments were separated from the carrier maltose-binding protein by digestion with human factor Xa or other proteases, and their structural integrity was confirmed by spectroscopic and calorimetric methods. The recombinant F1 and F4-5 exhibited fluorescence-detected melting transitions of the same magnitude and with the same midpoint (Tm) as their natural analogues prepared from Fib-1 by proteolysis. Differential scanning calorimetry experiments further demonstrated that these fragments are properly folded and have compact structures identical to the natural ones. Isolated rF4 melts at a much lower temperature than rF5 or the bimodular fragment rF4-5, indicating the loss of a stabilizing interaction between fingers 4 and 5. Comparison of fluorescence spectra of individual rF4 and rF5 with that of rF4-5 was also consistent with an interaction that affects the environment of Trp residue(s). rF2 also melts at a lower temperature than rF3 or rF2-3, suggesting a stabilizing interaction between the second and third fingers as well. When tested on fibrin-Sepharose, only the bimodular fragment rF4-5 was able to bind. All other fragments, including individual fingers 4 and 5, failed to bind. Thus, fibrin binding is not a common property of all fingers. The results indicate that a recognition site for fibrin is located within fingers 4 and 5. The interaction between these neighboring domains may play an important role in proper orientation of the residues forming this site.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
269
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
9539-46
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:8144540-Amino Acid Sequence, pubmed-meshheading:8144540-Base Sequence, pubmed-meshheading:8144540-Calorimetry, Differential Scanning, pubmed-meshheading:8144540-Chromatography, Affinity, pubmed-meshheading:8144540-Circular Dichroism, pubmed-meshheading:8144540-DNA Primers, pubmed-meshheading:8144540-Escherichia coli, pubmed-meshheading:8144540-Factor XIII, pubmed-meshheading:8144540-Fibrin, pubmed-meshheading:8144540-Fibronectins, pubmed-meshheading:8144540-Models, Structural, pubmed-meshheading:8144540-Molecular Sequence Data, pubmed-meshheading:8144540-Protein Denaturation, pubmed-meshheading:8144540-Protein Folding, pubmed-meshheading:8144540-Protein Structure, Secondary, pubmed-meshheading:8144540-Recombinant Proteins, pubmed-meshheading:8144540-Restriction Mapping, pubmed-meshheading:8144540-Thermodynamics
pubmed:year
1994
pubmed:articleTitle
The NH2-terminal fibrin-binding site of fibronectin is formed by interacting fourth and fifth finger domains. Studies with recombinant finger fragments expressed in Escherichia coli.
pubmed:affiliation
J. Holland Laboratory, American Red Cross, Rockville, Maryland 20855.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't