title: Identification of transglutaminase substrates from porcine nucleus pulposus as potential amplifiers in cross-linking cell scaffolds.
abstract: Nucleus pulposus from the porcine intervertebral disc was separated chromatographically to discover substrates of microbial transglutaminase. Highly purified proteins were prepared, among them type II collagen, the major protein of the nucleus pulposus. Determination of substrates was performed by transglutaminase-mediated incorporation of biotinylated probes displaying several glutamine and lysine donor proteins. Type II collagen was only labeled if smaller nucleus pulposus proteins were present. One of the modulating proteins was serotransferrin, a lysine donor substrate of bacterial transglutaminase. An additional substrate was the carboxy-terminal propeptide of type II collagen, chondrocalcin. Chondrocalcin, a regulator of type II collagen fibrillogenesis, occurs abundantly in juvenile cartilage and nucleus pulposus. Accordingly, the protein may be regarded as an excellent additive for the preparation of injectable stem cells in nucleus pulposus-like matrices cross-linked by microbial transglutaminase.
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Applied Sciences of Darmstadt, Schnittspahnstrasse 12, 64287 Darmstadt, Germany.
authors.forename: Elke
authors.initials: E
authors.lastname: Gebauer
authors.forename: Elke
authors.initials: E
authors.lastname: Goßla
authors.forename: Carolin
authors.initials: C
authors.lastname: Kwas
authors.forename: Denise
authors.initials: D
authors.lastname: Salzig
authors.forename: Alexandra
authors.initials: A
authors.lastname: Schmiermund
authors.forename: Peter
authors.initials: P
authors.lastname: Czermak
authors.forename: Hans-Lothar
authors.initials: HL
authors.lastname: Fuchsbauer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002356:Cartilage / Q000737:chemistry*; D024043:Collagen Type II / Q000737:chemistry / Q000302:isolation & purification*; D007403:Intervertebral Disc / Q000737:chemistry*; D008969:Molecular Sequence Data; D011485:Protein Binding; D013302:Streptomyces / Q000737:chemistry* / Q000201:enzymology; D013379:Substrate Specificity; D013552:Swine; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014168:Transferrin / Q000737:chemistry* / Q000302:isolation & purification; D011503:Transglutaminases / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D002135:Calcium-Binding Proteins; D024043:Collagen Type II; D014168:Transferrin; C040072:chondrocalcin; D011503:Transglutaminases