PMID- 10636927
OWN - NLM
STAT- MEDLINE
DCOM- 20000224
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 3
DP  - 2000 Jan 21
TI  - Initial steps in assembly of microfibrils. Formation of disulfide-cross-linked
      multimers containing fibrillin-1.
PG  - 2205-10
AB  - Fibrillins are the major constituents of extracellular microfibrils. How
      fibrillin molecules assemble into microfibrils is not known. Sequential
      extractions and pulse-chase labeling of organ cultures of embryonic chick aortae 
      revealed rapid formation of disulfide-cross-linked aggregates containing
      fibrillin-1. These results demonstrated that intermolecular disulfide bond
      formation is an initial step in the assembly process. To identify free cysteine
      residues available for intermolecular cross-linking, small recombinant peptides
      of fibrillin-1 harboring candidate cysteine residues were analyzed. Results
      revealed that the first four cysteine residues in the unique N terminus form
      intramolecular disulfide bonds. One cysteine residue (Cys(204)) in the first
      hybrid domain of fibrillin-1 was found to occur as a free thiol and is therefore 
      a good candidate for intermolecular disulfide bonding in initial steps of the
      assembly process. Furthermore, evidence indicated that the comparable cysteine
      residue in fibrillin-2 (Cys(233)) also occurs as a free thiol. These free
      cysteine residues in fibrillins are readily available for intermolecular
      disulfide bond formation, as determined by reaction with Ellman's reagent. In
      addition to these major results, the cleavage site of the fibrillin-1 signal
      peptide and the N-terminal sequence of monomeric authentic fibrillin-1 from
      conditioned fibroblast medium were determined.
FAU - Reinhardt, D P
AU  - Reinhardt DP
AD  - Shriners Hospital for Children, Oregon Health Sciences University, Portland,
      Oregon 97201, USA.
FAU - Gambee, J E
AU  - Gambee JE
FAU - Ono, R N
AU  - Ono RN
FAU - Bachinger, H P
AU  - Bachinger HP
FAU - Sakai, L Y
AU  - Sakai LY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Disulfides)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (FBN1 protein, human)
RN  - 0 (FBN2 protein, human)
RN  - 0 (Fibrillin-1)
RN  - 0 (Fibrillin-2)
RN  - 0 (Fibrillins)
RN  - 0 (Microfilament Proteins)
RN  - 0 (Recombinant Proteins)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Aorta/embryology
MH  - Chick Embryo
MH  - Cross-Linking Reagents/*metabolism
MH  - Culture Media, Serum-Free/metabolism
MH  - Cysteine/metabolism
MH  - Disulfides/*metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Extracellular Matrix Proteins/chemistry
MH  - Fibrillin-1
MH  - Fibrillin-2
MH  - Fibrillins
MH  - Fibroblasts/metabolism
MH  - Humans
MH  - Microfibrils/chemistry/*metabolism
MH  - Microfilament Proteins/chemistry/*metabolism
MH  - Molecular Sequence Data
MH  - Mutagenesis
MH  - Organ Culture Techniques
MH  - Recombinant Proteins/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Time Factors
EDAT- 2000/01/15 09:00
MHDA- 2000/02/26 09:00
CRDT- 2000/01/15 09:00
PHST- 2000/01/15 09:00 [pubmed]
PHST- 2000/02/26 09:00 [medline]
PHST- 2000/01/15 09:00 [entrez]
AID - 10.1074/jbc.275.3.2205 [doi]
AID - S0021-9258(18)31160-8 [pii]
PST - ppublish
SO  - J Biol Chem. 2000 Jan 21;275(3):2205-10. doi: 10.1074/jbc.275.3.2205.