HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ritty, T. M. Articles by Mecham, R. P. Search for Related Content PubMed PubMed Citation Articles by Ritty, T. M. Articles by Mecham, R. P. Social Bookmarking                      What's this?

J Biol Chem, Vol. 274, Issue 13, 8933-8940, March 26, 1999

Processing of the Fibrillin-1 Carboxyl-terminal Domain

Timothy M. Ritty, Thomas Broekelmann, Clarina Tisdale, Dianna M. Milewicz^§, and Robert P. Mecham

From the  Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110 and the ^§ Department of Internal Medicine, University of Texas-Houston Medical School, Houston, Texas 77030

To investigate the processing and general properties of the fibrillin-1 carboxyl-terminal domain, three protein expression constructs have been developed as follows: one without the domain, one with the domain, and one with a mutation near the putative proteolytic processing site. The constructs have been expressed in two eukaryotic model systems, baculoviral and CHO-K1. Post-translational modifications that normally occur in fibrillin-1, including glycosylation, signal peptide cleavage, and carboxyl-terminal processing, occur in the three constructs in both cell systems. Amino-terminal sequencing of secreted protein revealed leader sequence processing at two sites, a primary site between Gly-24/Ala-25 and a secondary site of Ala-27/Asn-28. Processing of the carboxyl-terminal domain could be observed by migration differences in SDS-polyacrylamide gel electrophoresis and was evident in both mammalian and insect cells. Immunological identification by Western blotting confirmed the loss of the expected region. The failure of both cell systems to process the mutant construct shows that the multi-basic sequence is the site of proteolytic processing. Cleavage of the fibrillin-1 carboxyl-terminal domain occurred intracellularly in CHO-K1 cells in an early secretory pathway compartment as demonstrated by studies with secretion blocking agents. This finding, taken with the multi-basic nature of the cleavage site and observed calcium sensitivity of cleavage, suggests that the processing enzyme is a secretory pathway resident furin-like protease.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				J. S. Weinbaum, T. J. Broekelmann, R. A. Pierce, C. C. Werneck, F. Segade, C. S. Craft, R. H. Knutsen, and R. P. Mecham Deficiency in Microfibril-associated Glycoprotein-1 Leads to Complex Phenotypes in Multiple Organ Systems J. Biol. Chem., September 12, 2008; 283(37): 25533 - 25543. [Abstract] [Full Text] [PDF]

				S. E. McGowan, A. J. Holmes, R. P. Mecham, and T. M. Ritty Arg-Gly-Asp-Containing Domains of Fibrillins-1 and -2 Distinctly Regulate Lung Fibroblast Migration Am. J. Respir. Cell Mol. Biol., April 1, 2008; 38(4): 435 - 445. [Abstract] [Full Text] [PDF]

				P N Robinson, E Arteaga-Solis, C Baldock, G Collod-Beroud, P Booms, A De Paepe, H C Dietz, G Guo, P A Handford, D P Judge, et al. The molecular genetics of Marfan syndrome and related disorders J. Med. Genet., October 1, 2006; 43(10): 769 - 787. [Abstract] [Full Text] [PDF]

				D. Hubmacher, K. Tiedemann, R. Bartels, J. Brinckmann, T. Vollbrandt, B. Batge, H. Notbohm, and D. P. Reinhardt Modification of the Structure and Function of Fibrillin-1 by Homocysteine Suggests a Potential Pathogenetic Mechanism in Homocystinuria J. Biol. Chem., October 14, 2005; 280(41): 34946 - 34955. [Abstract] [Full Text] [PDF]

				S. Hutchinson, A. Furger, D. Halliday, D. P. Judge, A. Jefferson, H. C. Dietz, H. Firth, and P. A. Handford Allelic variation in normal human FBN1 expression in a family with Marfan syndrome: a potential modifier of phenotype? Hum. Mol. Genet., September 15, 2003; 12(18): 2269 - 2276. [Abstract] [Full Text] [PDF]

				B. A. Kozel, H. Wachi, E. C. Davis, and R. P. Mecham Domains in Tropoelastin That Mediate Elastin Deposition in Vitro and in Vivo J. Biol. Chem., May 9, 2003; 278(20): 18491 - 18498. [Abstract] [Full Text] [PDF]

				G. Lin, K. Tiedemann, T. Vollbrandt, H. Peters, B. Batge, J. Brinckmann, and D. P. Reinhardt Homo- and Heterotypic Fibrillin-1 and -2 Interactions Constitute the Basis for the Assembly of Microfibrils J. Biol. Chem., December 20, 2002; 277(52): 50795 - 50804. [Abstract] [Full Text] [PDF]

				C. M. Kielty, M. J. Sherratt, and C. A. Shuttleworth Elastic fibres J. Cell Sci., July 15, 2002; 115(14): 2817 - 2828. [Abstract] [Full Text] [PDF]

				S. S. Chaudhry, J. Gazzard, C. Baldock, J. Dixon, M. J. Rock, G. C. Skinner, K. P. Steel, C. M. Kielty, and M. J. Dixon Mutation of the gene encoding fibrillin-2 results in syndactyly in mice Hum. Mol. Genet., April 1, 2001; 10(8): 835 - 843. [Abstract] [Full Text] [PDF]

				C. Baldock, A. J. Koster, U. Ziese, M. J. Rock, M. J. Sherratt, K. E. Kadler, C. A. Shuttleworth, and C. M. Kielty The Supramolecular Organization of Fibrillin-rich Microfibrils J. Cell Biol., March 5, 2001; 152(5): 1045 - 1056. [Abstract] [Full Text] [PDF]

				J. L ASHWORTH, C. M KIELTY, and D. McLEOD Fibrillin and the eye Br. J. Ophthalmol., November 1, 2000; 84(11): 1312 - 1317. [Full Text] [PDF]

				A. J. McGettrick, V. Knott, A. Willis, and P. A. Handford Molecular effects of calcium binding mutations in Marfan syndrome depend on domain context Hum. Mol. Genet., August 12, 2000; 9(13): 1987 - 1994. [Abstract] [Full Text] [PDF]

				B. C. Trask, T. M. Trask, T. Broekelmann, and R. P. Mecham The Microfibrillar Proteins MAGP-1 and Fibrillin-1 Form a Ternary Complex with the Chondroitin Sulfate Proteoglycan Decorin Mol. Biol. Cell, May 1, 2000; 11(5): 1499 - 1507. [Abstract] [Full Text]

				D. P. Reinhardt, J. E. Gambee, R. N. Ono, H. P. Bachinger, and L. Y. Sakai Initial Steps in Assembly of Microfibrils. FORMATION OF DISULFIDE-CROSS-LINKED MULTIMERS CONTAINING FIBRILLIN-1 J. Biol. Chem., January 21, 2000; 275(3): 2205 - 2210. [Abstract] [Full Text] [PDF]

				P. N Robinson and M. Godfrey The molecular genetics of Marfan syndrome and related microfibrillopathies J. Med. Genet., January 1, 2000; 37(1): 9 - 25. [Abstract] [Full Text]

				J. Ashworth, V Kelly, M. Rock, C. Shuttleworth, and C. Kielty Regulation of fibrillin carboxy-terminal furin processing by N-glycosylation, and association of amino- and carboxy-terminal sequences J. Cell Sci., January 11, 1999; 112(22): 4163 - 4171. [Abstract] [PDF]

				J. Ashworth, V Kelly, R Wilson, C. Shuttleworth, and C. Kielty Fibrillin assembly: dimer formation mediated by amino-terminal sequences J. Cell Sci., January 10, 1999; 112(20): 3549 - 3558. [Abstract] [PDF]

				T. M. Trask, B. C. Trask, T. M. Ritty, W. R. Abrams, J. Rosenbloom, and R. P. Mecham Interaction of Tropoelastin with the Amino-terminal Domains of Fibrillin-1 and Fibrillin-2 Suggests a Role for the Fibrillins in Elastic Fiber Assembly J. Biol. Chem., August 4, 2000; 275(32): 24400 - 24406. [Abstract] [Full Text] [PDF]