PMID- 10196183
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 16
DP  - 1999 Apr 16
TI  - Involucrin cross-linking by transglutaminase 1. Binding to membranes directs
      residue specificity.
PG  - 11013-21
AB  - The transglutaminase 1 (TGase 1) enzyme is essential for the assembly of the cell
      envelope barrier in stratified squamous epithelia. It is usually bound to
      membranes, but to date most studies with it have involved solution assays. Here
      we describe an in vitro model system for characterizing the function of TGase 1
      on the surface of synthetic lipid vesicles (SLV) of composition similar to
      eukaryote plasma membranes. Recombinant baculovirus-expressed human TGase 1
      readily binds to SLV and becomes active in cross-linking above 10 microM Ca2+, in
      comparison to above 100 microM in solution assays, suggesting that the membrane
      surface is important for enzyme function. Involucrin also binds to SLV containing
      12-18% phosphatidylserine and at Ca2+ concentrations above 1 microM. In reactions
      of involucrin with TGase 1 enzyme in solution, 80 of its 150 glutamines serve as 
      donor residues. However, on SLV carrying both involucrin and TGase 1, only five
      glutamines serve as donors, of which glutamine 496 was the most favored. As
      controls, there was no change in specificity toward the glutamines of other
      substrates used by free or SLV-bound TGase 1 enzyme. We propose a model in which 
      involucrin and TGase 1 bind to membranes shortly after expression in
      differentiating keratinocytes, but cross-linking begins only later as
      intracellular Ca2+ levels increase. Furthermore, the data suggest that the
      membrane surface regulates the steric interaction of TGase 1 with substrates such
      as involucrin to permit specific cross-linking for initiation of cell envelope
      barrier formation.
FAU - Nemes, Z
AU  - Nemes Z
AD  - Laboratory of Skin Biology, NIAMS, National Institutes of Health, Bethesda,
      Maryland 20892-2752, USA.
FAU - Marekov, L N
AU  - Marekov LN
FAU - Steinert, P M
AU  - Steinert PM
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA Primers)
RN  - 0 (Protein Precursors)
RN  - 0 (Recombinant Proteins)
RN  - 60108-77-2 (involucrin)
RN  - EC 2.3.2.13 (Transglutaminases)
RN  - EC 2.3.2.13 (transglutaminase 1)
RN  - SY7Q814VUP (Calcium)
RN  - TE7660XO1C (Glycine)
SB  - IM
MH  - Base Sequence
MH  - Calcium/metabolism
MH  - DNA Primers
MH  - Glycine/metabolism
MH  - Humans
MH  - Kinetics
MH  - Lipid Metabolism
MH  - Protein Precursors/*metabolism
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Solubility
MH  - Substrate Specificity
MH  - Transglutaminases/chemistry/*metabolism
EDAT- 1999/04/10 00:00
MHDA- 1999/04/10 00:01
CRDT- 1999/04/10 00:00
PHST- 1999/04/10 00:00 [pubmed]
PHST- 1999/04/10 00:01 [medline]
PHST- 1999/04/10 00:00 [entrez]
AID - 10.1074/jbc.274.16.11013 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Apr 16;274(16):11013-21. doi: 10.1074/jbc.274.16.11013.