# KERATINOCYTE TRANSGLUTAMINASE: ANCHORAGE AND CLONING

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA AT DAVIS · 1991 · $156,615

## Abstract

This proposal concerns elucidation of the primary structure of
keratinocyte transglutaminase and localization of posttranslational
fatty acid and phosphate modifications.  First, the complete
nucleotide sequence of the cDNA will be obtained.  Positive cDNA
clones have been selected in an oligo dT-primed lambda gt 11
library with a polyclonal antiserum raised to the purified enzyme.
With these probes, the library will be rescreened to identify
longer clones covering most of the message sequence.  As necessary,
new libraries will be prepared with random primers or by primer
extension using synthetic oligonucleotides.  Genomic clones will
then be selected and sequenced, including up to 2 kb of 5'-flanking
DNA.  To assist in planning the cDNA sequencing, the size of the
transglutaminase mRNA will be estimated by northern blot analysis.
Cell type specificity of expression of the message will be examined
as well as regulation of message levels in keratinocytes by
physiological agents known to modulate greatly the expression of
the enzyme (retinoids, hydrocortisone, calcium).  The
transglutaminase amino acid sequence will be deduced from the cDNA
nucleotide sequence to assist with determining the orientation of
the enzyme in the membrane and sites of posttranslational
modification.  An anchorage region remaining in the membrane after
trypsin release of an active 80 kd fragment will be purified and
the N-terminal amino acid sequence obtained.  Peptides containing
the fatty acid and phosphate modifications will be purified
(primarily by HPLC techniques) and sequenced.  In addition, the
degree of phosphorylation of the transglutaminase will be
quantitated.  The results of this work will provide a basis for
elucidating (i) the role of keratinocyte transglutaminase in cross-
linked envelope formation from a molecular perspective (orientation
in the membrane, interaction with substrate proteins), (ii)
physiological regulation of the enzyme expression by molecular
biological techniques, (iii) molecular mechanisms of keratinocyte
reprogramming in pathological conditions, and (iv) evolution of
transglutaminases.

## Key facts

- **NIH application ID:** 3155527
- **Project number:** 5R01AR027130-11
- **Recipient organization:** UNIVERSITY OF CALIFORNIA AT DAVIS
- **SAM.gov UEI:** TX2DAGQPENZ5
- **Principal Investigator:** ROBERT H RICE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $156,615
- **Award type:** 5
- **Project period:** 1989-09-30 → 1992-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3155527

## Citation

> US National Institutes of Health, RePORTER application 3155527, KERATINOCYTE TRANSGLUTAMINASE: ANCHORAGE AND CLONING (5R01AR027130-11). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3155527. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*