Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. 1976 Dec; 73(12): 4479–4481.
PMCID: PMC431507

Role of the intrinsic transglutaminase in the Ca2+-mediated crosslinking of erythrocyte proteins.


Transamidase (i.e., "transglutaminase") activity of human erythrocytes, lysed by a single freezing and thawing to 37 degrees, was measured by a method of incorporating [14C]putrescine into N,N'-dimethylcasein. In the absence of added calcium ions, virtually no enzyme activity could be detected. An increase in concentration of the cation to about 0.5 mM, however, turned on the enzyme to appreciable levels of activity. Simultaneously, Ca2+ produced formation of high molecular weight, nondisulfied bonded protein polymers either directly in the lysate or in fresh cells when the cation was added together with the A23187 ionophore. The polymers could be readily identified in the isolated cell ghosts by means of disc gel electrophoresis. If the Ca2+-promoted formation of polymers was allowed to take place in the presence of 14C-putrescine, then this tracer became incorporated into the polymeric material. The incorporation indicated that polymerization occurred through gamma-glutamyl-epsilon-lysine bridtes. It is suggested that the intrinsic transamidase mediates protein crosslinking of the erythrocyte membrane whenever there is an increase in intracellular Ca2+ concentration. The presence of suitable transglutaminase substrates, e.g. histamine, inhibited crosslinking when the cells were incubated with Ca2+ and ionophore.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (703K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Dutton A, Singer SJ. Crosslinking and labeling of membrane proteins by transglutaminase-catalyzed reactions. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2568–2571. [PMC free article] [PubMed]
  • Lorand L, Shishido R, Parameswaran KN, Steck TL. Modification of human erythrocyte ghosts with transglutaminase. Biochem Biophys Res Commun. 1975 Dec 1;67(3):1158–1166. [PubMed]
  • Fairbanks G, Steck TL, Wallach DF. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. [PubMed]
  • Lin Y, Means GE, Feeney RE. The action of proteolytic enzymes on N,N-dimethyl proteins. Basis for a microassay for proteolytic enzymes. J Biol Chem. 1969 Feb 10;244(3):789–793. [PubMed]
  • Lorand L, Campbell-Wilkes LK, Cooperstein L. A filter paper assay for transamidating enzymes using radioactive amine substrates. Anal Biochem. 1972 Dec;50(2):623–631. [PubMed]
  • Steck TL, Yu J. Selective solubilization of proteins from red blood cell membranes by protein perturbants. J Supramol Struct. 1973;1(3):220–232. [PubMed]
  • Tishler PV, Epstein CJ. A convenient method of preparing polyacrylamide gels for liquid scintillation spectrometry. Anal Biochem. 1968 Jan;22(1):89–98. [PubMed]
  • Carraway KL, Triplett RB, Anderson DR. Calcium-promoted aggregation of erythrocyte membrane proteins. Biochim Biophys Acta. 1975 Feb 27;379(2):571–581. [PubMed]
  • Lorand L, Chenoweth D, Gray A. Titration of the acceptor cross-linking sites in fibrin. Ann N Y Acad Sci. 1972 Dec 8;202:155–171. [PubMed]
  • Eaton JW, Skelton TD, Swofford HS, Kolpin CE, Jacob HS. Elevated erythrocyte calcium in sickle cell disease. Nature. 1973 Nov 9;246(5428):105–106. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • Nucleotide
    Published Nucleotide sequences
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...